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'description' => '<p style="text-align: justify;">The <strong>Auto Universal Plant ChIP-seq</strong> kit offers the convenience of extracting plant chromatin from a wide variety of plants including Arabidopsis, maize, rice, tomato and poplar and has been validated for the <strong>IP-Star® automated system</strong>. This complete kit has been specifically optimized for <strong>plant chromatin extraction</strong> and includes reagents for chromatin preparation, immunoprecipitation, plant-specific control primer pairs, control antibody, and DNA purification.</p>',
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<li>Validated for the high throughput <strong>IP-Star® Automated System</strong></li>
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<h3>Successful ChIP-seq experiments for a variety of plants</h3>
<div class="row">
<div class="small-6 columns"><center>Arabidopsis</center><center><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></center>
<p><small><strong>Figure 1.</strong> ChIP-seq was performed on Arabidopsis thaliana (Col-0) seedlings using our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng (green), 500 pg (orange) and 100 pg (red) IP'd DNA and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a public dataset (NCBI GEO Dataset GSM1193621) that we used as an external reference. Enrichments along a wide region of chromosome 5 are uniform regardless of the starting material amount for the preparation of the library.</small></p>
</div>
<div class="small-6 columns"><center>Poplar</center><center><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></center>
<p><small><strong>Figure 3.</strong> ChIP-seq was performed on Populus trichocarpa stem differenciating xylem using the Premium H3K4me3 ChIP-seq grade antibody. Libraries were prepared with the <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the input and is considered as the background enrichment. The profile in red represents enrichments along a wide region of scaffold 18. Using the same scale, the peaks of the immunoprecipitated samples are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center>Tomato</center><center><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></center>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our Premium H3K4me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
</div>
<div class="small-6 columns"><center>Maize</center><center><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></center>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our Premium H3K27me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<table style="width: 856px;">
<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
</td>
<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
</td>
<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
</td>
</tr>
<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9-14ac-polyclonal-antibody-classic-sample-size-10-mg">H3K9/14ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27ac-polyclonal-antibody-premium-sample-size-10-ug">H3K27ac polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
</tbody>
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<li>Validated for the high throughput <strong>IP-Star® Automated System</strong></li>
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<h3>Successful ChIP-seq experiments for a variety of plants</h3>
<div class="row">
<div class="small-6 columns"><center>Arabidopsis</center><center><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></center>
<p><small><strong>Figure 1.</strong> ChIP-seq was performed on Arabidopsis thaliana (Col-0) seedlings using our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng (green), 500 pg (orange) and 100 pg (red) IP'd DNA and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a public dataset (NCBI GEO Dataset GSM1193621) that we used as an external reference. Enrichments along a wide region of chromosome 5 are uniform regardless of the starting material amount for the preparation of the library.</small></p>
</div>
<div class="small-6 columns"><center>Poplar</center><center><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></center>
<p><small><strong>Figure 3.</strong> ChIP-seq was performed on Populus trichocarpa stem differenciating xylem using the Premium H3K4me3 ChIP-seq grade antibody. Libraries were prepared with the <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the input and is considered as the background enrichment. The profile in red represents enrichments along a wide region of scaffold 18. Using the same scale, the peaks of the immunoprecipitated samples are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center>Tomato</center><center><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></center>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our Premium H3K4me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
</div>
<div class="small-6 columns"><center>Maize</center><center><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></center>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our Premium H3K27me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<table style="width: 856px;">
<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
</td>
<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
</td>
<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
</td>
</tr>
<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9-14ac-polyclonal-antibody-classic-sample-size-10-mg">H3K9/14ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
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<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
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<td style="width: 224px;"></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
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<tr>
<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
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<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
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<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
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<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the trimethylated lysine 4</strong> (<strong>H3K4me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4me3</strong><br />ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K4me3 (cat. No. C15410003) and optimized PCR primer pairs for qPCR. ChIP was performed with the iDeal ChIP-seq kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p></p>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2a-ChIP-seq.jpg" width="800" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2b-ChIP-seq.jpg" width="800" /></center><center>C.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2c-ChIP-seq.jpg" width="800" /></center><center>D.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2d-ChIP-seq.jpg" width="800" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K4me3</strong><br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of the Diagenode antibody against H3K4me3 (cat. No. C15410003) as described above. The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 600 kb region of the X-chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes, respectively (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</small></p>
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<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-cuttag-a.png" width="800" /></center></div>
<div class="small-12 columns"><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-cuttag-b.png" width="800" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K4me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 0.5 µg of the Diagenode antibody against H3K4me3 (cat. No. C15410003) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the ACTB gene on chromosome 7 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig3-ELISA.jpg" width="350" /></center><center></center><center></center><center></center><center></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody against H3K4me3 (cat. No. C15410003). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:11,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig4-DB.jpg" /></div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K4me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K4me3 (cat. No. C15410003), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 5A shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig5-WB.jpg" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K4me3</strong><br />Western blot was performed on whole cell extracts (40 µg, lane 1) from HeLa cells, and on 1 µg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K4me3 (cat. No. C15410003). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig6-if.jpg" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K4me3</strong><br />HeLa cells were stained with the Diagenode antibody against H3K4me3 (cat. No. C15410003) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. The cells were immunofluorescently labelled with the H3K4me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (middle), which specifically labels DNA. The right picture shows a merge of both stainings.</small></p>
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'meta_description' => 'H3K4me3 (Histone H3 trimethylated at lysine 4) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array. Batch-specific data available on the website. Sample size available.',
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'id' => '2264',
'antibody_id' => '121',
'name' => 'H3K9me3 Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone<strong> H3 containing the trimethylated lysine 9</strong> (<strong>H3K9me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig1.png" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9me3</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K9me3 (cat. No. C15410193) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, 2, and 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for the heterochromatin marker Sat2 and for the ZNF510 gene, used as positive controls, and for the promoters of the active EIF4A2 and GAPDH genes, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2a.png" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2b.png" width="700" /></center><center>C.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2c.png" width="700" /></center><center>D.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2d.png" width="700" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K9me3</strong><br />ChIP was performed with 0.5 µg of the Diagenode antibody against H3K9me3 (cat. No. C15410193) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2A shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. The arrows indicate two satellite repeat regions which exhibit a stronger signal. Figures 2B, 2C and 2D show the enrichment along the ZNF510 positive control target and at the H19 and KCNQ1 imprinted genes.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-CT-Fig3a.png" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-CT-Fig3b.png" width="700" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K9me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K9me3 (cat. No. C15410193) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in a genomic regions on chromosome 1 containing several ZNF repeat genes and in a genomic region surrounding the KCNQ1 imprinting control gene on chromosome 11 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-Elisa-Fig4.png" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human H3K9me3 (cat. No. C15410193) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:87,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-DB-Fig5.png" /></center></div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K9me3</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9me3 (cat. No. C15410193) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-WB-Fig6.png" /></center></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K9me3</strong><br />Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K9me3 (cat. No. C15410193). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-IF-Fig7.png" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K9me3</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9me3 (cat. No. C15410193) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me3 antibody (middle) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of both stainings is shown on the right.</small></p>
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'meta_title' => 'H3K9me3 Antibody - ChIP-seq Grade (C15410193) | Diagenode',
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'meta_description' => 'H3K9me3 (Histone H3 trimethylated at lysine 9) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array assay. Batch-specific data available on the website. Sample size available.',
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'name' => 'H3pan Antibody',
'description' => '<p><span>This antibody has been raised in rabbit against two KLH-conjugated synthetic peptides containing an unmodified sequence from the central part and from the C-terminus of <strong>histone H3</strong>, respectively.</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-chip.jpg" alt="H3pan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3pan</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3pan (Cat. No. C15310135) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 μl per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive MYOD1 and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-elisa.jpg" alt="H3pan Antibody ELISA validation" caption="false" width="288" height="217" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3pan (Cat. No. C15310135). The plates were coated with the peptides used for immunization. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be >1:1,000,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-wb.jpg" alt="H3pan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against H3pan</strong><br />Whole cell extracts from HeLa cells (25 μg) were analysed by Western blot using the Diagenode antibody against H3pan (Cat. No. C15310135) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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'meta_description' => 'H3pan (Histone H3) Polyclonal Antibody validated in ChIP-qPCR, WB and ELISA. Batch-specific data available on the website. Sample size available.',
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'name' => 'H3K27me3 Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the trimethylated lysine 27</strong> (<strong>H3K27me3</strong>), using a KLH-conjugated synthetic peptide.</p>',
'label1' => 'Validation Data',
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<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig1.png" alt="H3K27me3 Antibody ChIP Grade" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2.png" alt="H3K27me3 Antibody for ChIP" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K27me3 (Cat. No. C15410195) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control.</small></p>
<p><small><strong>Figure 1A.</strong> Quantitative PCR was performed with primers specific for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive TSH2B and MYT1 genes, used as positive controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
<p><small><strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me3, H3K9me3 and H3K36me3 modifications and the unmodified H3K27 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H3K27me3 modification.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2a.png" alt="H3K27me3 Antibody ChIP-seq Grade" /></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-12 columns">
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2b.png" alt="H3K27me3 Antibody for ChIP-seq" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2c.png" alt="H3K27me3 Antibody for ChIP-seq assay" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2d.png" alt="H3K27me3 Antibody validated in ChIP-seq" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 µg of the Diagenode antibody against H3K27me3 (Cat. No. C15410195) as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment in genomic regions of chromosome 6 and 20, surrounding the TSH2B and MYT1 positive control genes (fig 2A and 2B, respectively), and in two genomic regions of chromosome 1 and X (figure 2C and D).</small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-CUTTAG-Fig3A.png" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-CUTTAG-Fig3B.png" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K27me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K27me3 (cat. No. C15410195) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions on chromosome and 13 and 20 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-ELISA-Fig4.png" alt="H3K27me3 Antibody ELISA Validation " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me3 (Cat. No. C15410195). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:3,000.</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-DB-Fig5a.png" alt="H3K27me3 Antibody Dot Blot Validation " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K27me3</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3 (Cat. No. C15410195) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 5 shows a high specificity of the antibody for the modification of interest. Please note that the antibody also recognizes the modification if S28 is phosphorylated.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-WB-Fig6.png" alt="H3K27me3 Antibody validated in Western Blot" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K27me3 (cat. No. C15410195) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-IF-Fig7.png" alt="H3K27me3 Antibody validated for Immunofluorescence" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K27me3</strong><br />Human HeLa cells were stained with the Diagenode antibody against H3K27me3 (Cat. No. C15410195) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'slug' => 'h3k27me3-polyclonal-antibody-premium-50-mg-27-ml',
'meta_title' => 'H3K27me3 Antibody - ChIP-seq Grade (C15410195) | Diagenode',
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'meta_description' => 'H3K27me3 (Histone H3 trimethylated at lysine 27) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array assay. Batch-specific data available on the website. Sample size available.',
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'name' => 'Chromatin EasyShear Kit for Plant ',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/Chromatin_Shearing_Optimization-Universal_Plant.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p style="text-align: justify;"><span>Previous name of the kit: Chromatin Shearing Optimization Kit (Universal Plant ChIP-seq kit)<br /></span></p>
<p style="text-align: justify;"><span>The first critical step of a successful ChIP experiment is the best preparation of sheared chromatin. This <strong>Chromatin EasyShear Kit</strong> is designed to be used in conjunction with the <strong>Universal Plant ChIP-seq kit</strong> and contains the right level of <strong>detergent</strong> for extraction of highest quality plant chromatin for ChIP. In addition, the signature</span><span> crosslinking containers of this kit provide a simple and reliable method for fixation. The content of this kit is enough to perform 12 chromatin extractions.<br /></span></p>
<p style="text-align: justify;"><span>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
<p style="text-align: justify;"><span>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></span></p>',
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'slug' => 'chromatin-shearing-plant-chip-seq-kit',
'meta_title' => 'Chromatin Shearing Optimization Kit (Universal Plant ChIP-seq kit) | Diagenode',
'meta_keywords' => 'chromatin shearing, plant epigenetics, plant ChIP, plant ChIP-seq, Arabidopsis, maize, rice, tomato, poplar',
'meta_description' => 'Chromatin Shearing Optimization Kit designed to be used in conjunction with the Universal Plant ChIP-seq kit, providing high quality plant chromatin for ChIP',
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<p style="text-align: justify;">The <strong>Universal Plant ChIP-seq kit</strong> offers the convenience of extracting plant chromatin from a wide variety of plants including Arabidopsis, maize, rice, tomato and poplar. This complete kit has been specifically optimized for <strong>plant chromatin extraction</strong> and includes reagents for chromatin preparation, immunoprecipitation, plant-specific control primer pairs, control antibody, and DNA purification.</p>',
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<li><strong>Universal compatiblity</strong> with a wide variety of plant species</li>
<li>Optimized and <strong>complete kit</strong> for start-to-finish plant ChIP</li>
<li>Includes <strong>plant-specific control</strong> primers and control antibody<strong></strong></li>
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<h3>Successful ChIP-seq experiments for a variety of plants</h3>
<div class="row">
<div class="small-6 columns">
<h4 class="text-center">Arabidopsis</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG1"> <img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A-small.jpg" /> </a></p>
<div id="IMG1" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 1.</strong> ChIP-seq was performed on Arabidopsis thaliana (Col-0) seedlings using our <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng (green), 500 pg (orange) and 100 pg (red) IP'd DNA and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a public dataset (NCBI GEO Dataset GSM1193621) that we used as an external reference. Enrichments along a wide region of chromosome 5 are uniform regardless of the starting material amount for the preparation of the library.</small></p>
</div>
<div class="small-6 columns">
<h4 class="text-center">Poplar</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG2"><img src="https://www.diagenode.com/img/landing-pages/poplar-small.jpg" /> </a></p>
<div id="IMG2" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 3.</strong> ChIP-seq was performed on Populus trichocarpa stem differenciating xylem using the <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with the <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the input and is considered as the background enrichment. The profile in red represents enrichments along a wide region of scaffold 18. Using the same scale, the peaks of the immunoprecipitated samples are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<h4 class="text-center">Tomato</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG3"> <img src="https://www.diagenode.com/img/landing-pages/tomtato-small.jpg" /> </a></p>
<div id="IMG3" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
</div>
<div class="small-6 columns">
<h4 class="text-center">Maize</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG4"> <img src="https://www.diagenode.com/img/landing-pages/maize-small.jpg" /> </a></p>
<div id="IMG4" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our <a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">Premium H3K27me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<p><strong> </strong></p>
<table style="width: 856px;">
<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
</td>
<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
</td>
<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
</td>
</tr>
<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9-14ac-polyclonal-antibody-classic-sample-size-10-mg">H3K9/14ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27ac-polyclonal-antibody-premium-sample-size-10-ug">H3K27ac polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
</tbody>
</table>
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'meta_title' => 'Universal Plant ChIP-seq kit | Diagenode',
'meta_keywords' => 'plant epigenetics, plant ChIP, plant ChIP-seq, Arabidopsis, maize, rice, tomato, poplar',
'meta_description' => 'Optimized extraction of plant chromatin from Arabidopsis,maize,rice,tomato,poplar.Complete ChIP kit including plant-specific control primer pairs and antibody',
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<p>Diagenode’s<span> </span><b>IPure</b><b><span> </span>kit<span> </span></b>is the only DNA purification kit using magnetic beads, that is specifically optimized for extracting DNA from<span> </span><b>ChIP</b><b>,<span> </span></b><b>MeDIP</b><span> </span>and<span> </span><b>CUT&Tag</b>. The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification. This simple and straightforward protocol delivers pure DNA ready for any downstream application (e.g. next generation sequencing). Comparing to phenol-chloroform extraction, the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples.</p>
<center>
<h4>High DNA recovery after purification of ChIP samples using IPure technology</h4>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-chromatin-function.png" width="500" /></center>
<p></p>
<p><small>ChIP assays were performed using different amounts of U2OS cells and the H3K9me3 antibody (Cat. No.<span> </span><span>C15410056</span>; 2 g/IP). <span>The purified DNA was eluted in 50 µl of water and quantified with a Nanodrop.</span></small></p>
<p></p>
<p><strong>Benefits of the IPure kit:</strong></p>
<ul>
<li style="text-align: left;">Provides pure DNA for any downstream application (e. g. Next generation sequencing)</li>
<li style="text-align: left;">Non-toxic</li>
<li style="text-align: left;">Fast & easy to use</li>
<li style="text-align: left;">Optimized for DNA purification after ChIP, MeDIP and CUT&Tag</li>
<li style="text-align: left;">Compatible with automation</li>
<li style="text-align: left;">Validated on the IP-Star Compact</li>
</ul>
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'info1' => '<h2>IPure after ChIP</h2>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-A.png" alt="ChIP-seq figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-B.png" alt="ChIP-seq figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-C.png" alt="ChIP-seq figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small><strong>Figure 1.</strong> Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors (containing the IPure module for DNA purification) and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.</small></p>
<p></p>
<h2>IPure after CUT&Tag</h2>
<p>Successful CUT&Tag results showing a low background with high region-specific enrichment has been generated using 50.000 of K562 cells, 1 µg of H3K4me3 or H3K27me3 antibody (Diagenode, C15410003 or C15410069, respectively) and proteinA-Tn5 (1:250) (Diagenode, C01070001). 1 µg of IgG (C15410206) was used as negative control. Samples were purified using the IPure kit v2 or phenol-chloroform purification. The below figures present the comparison of two purification methods.</p>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-fig2.png" style="display: block; margin-left: auto; margin-right: auto;" width="400" /></center><center>
<p style="text-align: center;"><small><strong>Figure 2.</strong> Heatmap 3kb upstream and downstream of the TSS for H3K4me3</small></p>
</center>
<p></p>
<p><img src="https://www.diagenode.com/img/product/kits/ipure-fig3.png" style="display: block; margin-left: auto; margin-right: auto;" width="600" /></p>
<p></p>
<center><small><strong>Figure 3.</strong> Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using Diagenode’s pA-Tn5 transposase (Cat. No. C01070002), H3K27me3 antibody (Cat. No. C15410069) and IPure kit v2 vs phenol chloroform purification (PC).</small></center>
<p></p>
<p></p>
<h2>IPure after MeDIP</h2>
<center><img src="https://www.diagenode.com/img/product/kits/magmedip-seq-figure_multi3.jpg" alt="medip sequencing coverage" width="600" /></center><center></center><center>
<p></p>
<small><strong>Figure 4.</strong> Consistent coverage and methylation detection from different starting amounts of DNA with the Diagenode MagMeDIP-seq Package (including the Ipure kit for DNA purification). Samples containing decreasing starting amounts of DNA (from the top down: 1000 ng (red), 250 ng (blue), 100 ng (green)) originating from human blood were prepared, revealing a consistent coverage profile for the three different starting amounts, which enables reproducible methylation detection. The CpG islands (CGIs) (marked by yellow boxes in the bottom track) are predominantly unmethylated in the human genome, and as expected, we see a depletion of reads at and around CGIs.</small></center>
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<p><img src="https://www.diagenode.com/img/product/kits/workflow-ipure-cuttag.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h3><strong>Workflow description</strong></h3>
<h5><strong>IPure after ChIP</strong></h5>
<p><strong>Step 1:</strong> Chromatin is decrosslinked and eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added.<br /> <strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet.<br /> <strong>Step 3:</strong> Proteins and remaining buffer are washed away.<br /> <strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after MeDIP</strong></h5>
<p><strong>Step 1:</strong> DNA is eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Remaining buffer are washed away.<br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after CUT&Tag</strong></h5>
<p><strong>Step 1:</strong> pA-Tn5 is inactivated and DNA released from the cells. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Proteins and remaining buffer are washed away. <br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).</p>
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'description' => '<div class="row">
<div class="large-12 columns">Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on a genome-wide scale. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</div>
<div class="large-12 columns"></div>
<h5 class="large-12 columns"><strong></strong></h5>
<h5 class="large-12 columns"><strong>The ChIP-seq workflow</strong></h5>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/chip-seq-diagram.png" /></div>
<div class="large-12 columns"><br />
<ol>
<li class="large-12 columns"><strong>Chromatin preparation: </strong>Crosslink chromatin-bound proteins (histones or transcription factors) to DNA followed by cell lysis.</li>
<li class="large-12 columns"><strong>Chromatin shearing:</strong> Fragment chromatin by sonication to desired fragment size (100-500 bp)</li>
<li class="large-12 columns"><strong>Chromatin IP</strong>: Capture protein-DNA complexes with <strong><a href="../categories/chip-seq-grade-antibodies">specific ChIP-seq grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: Reverse cross-links, elute, and purify </li>
<li class="large-12 columns"><strong>NGS Library Preparation</strong>: Ligate adapters and amplify IP'd material</li>
<li class="large-12 columns"><strong>Bioinformatic analysis</strong>: Perform r<span style="font-weight: 400;">ead filtering and trimming</span>, r<span style="font-weight: 400;">ead specific alignment, enrichment specific peak calling, QC metrics, multi-sample cross-comparison etc. </span></li>
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</div>
<div class="row" style="margin-top: 32px;">
<div class="small-12 medium-10 large-9 small-centered columns">
<div class="radius panel" style="background-color: #fff;">
<h3 class="text-center" style="color: #b21329;">Need guidance?</h3>
<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
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<p class="text-justify">Chromatin Immunoprecipitation (ChIP) coupled with quantitative PCR can be used to investigate protein-DNA interaction at known genomic binding sites. if sites are not known, qPCR primers can also be designed against potential regulatory regions such as promoters. ChIP-qPCR is advantageous in studies that focus on specific genes and potential regulatory regions across differing experimental conditions as the cost of performing real-time PCR is minimal. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</p>
<p class="text-justify"><strong>The ChIP-qPCR workflow</strong></p>
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<li class="large-12 columns"><strong>Chromatin preparation: </strong>cell fixation (cross-linking) of chromatin-bound proteins such as histones or transcription factors to DNA followed by cell lysis.</li>
<li class="large-12 columns"><strong>Chromatin shearing: </strong>fragmentation of chromatin<strong> </strong>by sonication down to desired fragment size (100-500 bp)</li>
<li class="large-12 columns"><strong>Chromatin IP</strong>: protein-DNA complexe capture using<strong> <a href="https://www.diagenode.com/en/categories/chip-grade-antibodies">specific ChIP-grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: chromatin reverse cross-linking and elution followed by purification<strong> </strong></li>
<li class="large-12 columns"><strong>qPCR and analysis</strong>: using previously designed primers to amplify IP'd material at specific loci</li>
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<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
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<p><img src="https://www.diagenode.com/img/areas/plant.jpg" /></p>
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<h2>Epigenetic Regulation in Plants</h2>
<p>Plants utilize a number of gene regulation mechanisms to ensure proper development, function, growth, and survival under different environmental conditions. Plants depend on changes in gene expression to respond to environmental stimuli, in which the full repertoire of histone modifications, DNA methylation, and small ncRNAs play an important role in epigenetic regulation.</p>
<p>Studying the epigenetics of model plants such as Arabidopsis thaliana have allowed researchers to understand pathways that maintain chromatin modifications as well as the mapping of modifications such as DNA methylation on a genome-wide scale. Small RNAs have also been implicated in playing a role in the distribution of chromatin modifications, and RNA may also play a role in the complex epigenetic interactions that occur between homologous sequences (Moazed et al, 2009). In the future, by understanding epigenetic control, researchers can uncover the research necessary to improve plant growth, yields, and transformation efficiency especially in the face of climate change and other environmental factors.</p>
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<h3 style="font-weight: 100; margin-top: 0;">Chromatin</h3>
<p>Chromatin consists of nucleosomes formed by a complex of histone proteins and DNA, which allows the packaging of DNA into the nucleus. The less condensed euchromatin represents transcriptionally active regions, while heterochromatin is usually inactive (Vaillant and Paszkowski, 2007). Chromatin state is known to be influenced by both DNA methylation and histone modifications which in turn impact gene expression and the structure of chromosomes. In a recent study, the role of chromatin modifications during plant reproduction elucidated 3-dimensional chromosome reorganization mediated by histones and DNA methylation (Dukowic-Schulze et al. 2017). In addition, gibberellins have been shown in increasing the level of histone acetylation, which affects regions of chromatin involved in maize seed germination (Zheng et al. 2017). Another study reports a novel function of a tomato histone deacetylase gene in the regulation of fruit ripening (Guo et al. 2017).</p>
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<p>In addition, multigene families encode transcription factors, with members found throughout the genome or clustered on the same chromosome. Numerous DNA binding proteins that interact with plant promoters have been identified -- some are similar to well-characterized transcription factors in animals or yeast, while others are unique to plants. For example, diverse members of the subfamily X of the plant-specific ethylene response factor (ERF) transcription factors coordinate stress signaling with wound repair activation. Tissue repair is also enhanced through a protein complex of ERF and GRAS TFs (Heyman et. al,.2018). A compilation of known plant transcription factors can be found in the plant transcription factor database at http://plntfdb.bio.uni-potsdam.de/v3.0/.</p>
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<p><img src="https://www.diagenode.com/img/areas/rna-strand.jpg" /></p>
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<h3 style="font-weight: 100; margin-top: 0;">RNA</h3>
<p>Recent research shows that a number of classes of small RNAs are key epigenetic regulators. In many cases, small RNAs have been implicated in DNA methylation and chromatin modification (Meyer, 2015). In addition, the role of small RNAs has been implicated in plant stress tolerance (Kumar et al., 2017). López-Galiano et al also provided insight into a coordinated function of a miRNA gene and histone modifications in regulating the expression of a WRKY transcription factor in response to stress.</p>
<p>RNA interference (RNAi) is another epigenetic mechanism that leads to small RNA generation, which mediates gene silencing at the post-transcriptional level. RNAi technology has immense potential for plant disease resistance.</p>
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<h3 style="font-weight: 100; margin-top: 0;">DNA methylation</h3>
<p>Plants, unlike animals, have three sites that can be methylated G, CHG (H can be A, C, T), and CHH (Law and Jacobsen, 2010). DNA methylation has attracted particular interest. In Arabidopsis, one-third of methylated genes occur in transcribed regions, and 5% of genes are methylated in promoter regions, suggesting that many of these are epigenetically regulated. (Zhang et al., 2006).</p>
<p>There are thousands of differentially methylated regions (DMRs) that influence phenotype by influencing gene expression. The analysis of epigenetic recombinant inbred line (epiRIL) plants from Arabidopsis points to the evidence of the influence of DMRs. An epiRIL results from crossing two genetically identical plants with differing DNA methylation levels (with one parent as a homozygous mutant for an essential DNA methylation maintenance gene). The offspring of these plants have similar genomes that vary only in methylation levels. Many traits have been studied using epiRILs -- flowering time, plant height, and response to abiotic stress, some of which have now been mapped to DMRs (Zhang et al. 2018)</p>
<p>Regulation by DNA methylation has been shown to be important in many aspects of plant development and response such as vernalization, hybrid vigor, and self-incompatibility (Itabashi et al. 2017). For example, vernalization treatments have shown reduced DNA methylation and subsequent initiation of flowering (Burn et al., 1993). Stress can also influence DNA methylation in plants as a response to environmental stimuli. (Steward et al., 2002; Song et al., 2012). A high degree of DNA methylation has also suggested the role in the improvement of plant fitness under different environmental conditions (Saéz-Laguna et al., 2014). In addition, methylation can affect normal fruit and hypomethylation predicts homeotic transformation and loss of fruit yield (Ong-Abdullah et al., 2015)</p>
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<p>DNA demethylation has also been implied in various aspects of plant development including pollen tube formation, embryogenesis, fruit ripening, stomatal development, and nodule formation ( Li et al. 2017). Demethylation of rice genomic DNA caused an altered pattern of gene expression, inducing dwarf plants (Sano et al., 1990).</p>
<p>Epigenetic modifications contribute to the stability and survival of the plants and their ability to adapt in different environmental conditions.</p>
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<h3>Diagenode products for your epigenomics research in plants</h3>
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<h3 class="text-center"><a href="https://www.diagenode.com/en/categories/chromatin-function">Chromatin analysis</a></h3>
<center><a href="https://www.diagenode.com/en/categories/chromatin-function"><img src="https://www.diagenode.com/img/cancer/chromatin-icon.png" /></a></center>
<p class="text-left">Understand the role of chromatin in plant function and development</p>
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<li><a href="https://www.diagenode.com/en/categories/chromatin-function">Learn about our chromatin analysis products</a></li>
<li><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns"> Learn about the Universal Plant ChIP Kit</a></li>
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<h3 class="text-center"><a href="https://www.diagenode.com/en/categories/dna-methylation" style="color: #30415c;">DNA methylation</a></h3>
<center><a href="https://www.diagenode.com/en/categories/dna-methylation"><img src="https://www.diagenode.com/img/cancer/dna-icon.png" /></a></center>
<p class="text-left">DNA methylation and demethylation and the effects on plant response and function</p>
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<li><a href="https://www.diagenode.com/en/categories/dna-methylation">Discover DNA methylation analysis solutions at any resolution</a></li>
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<h3 class="text-center"><span class="darkgrey">Non-coding RNAs</span></h3>
<center><img src="https://www.diagenode.com/img/cancer/non-coding-icon.png" /></center>
<p class="text-left">Discover noncoding RNAs in the regulation of gene expression in plants</p>
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<li><a href="https://www.diagenode.com/en/categories/Library-preparation-for-RNA-seq">Library prep for RNA-seq studies for ncRNAs</a></li>
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<h3>References</h3>
<p><small> Burn, J. et al (1993). DNA methylation, vernalization, and the initiation of flowering. Proc. Natl. Acad. Sci. U.S.A. 90, 287–291. doi: 10.1006/scdb.1996.0055 </small></p>
<p><small> Dukowic-Schulze S, Liu C, Chen C (2017) Not just gene expression: 3D implications of chromatin modifications during sexual plant reproduction. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2222-0</small></p>
<p><small> Guo J et al (2017) A histone deacetylase gene, SlHDA3, acts as a negative regulator of fruit ripening and carotenoid accumulation. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2211-3</small></p>
<p><small> Heyman J, et.al (2018) Journal of Cell Science Emerging role of the plant ERF transcription factors in coordinating wound defense responses and repair doi: 10.1242/jcs.208215</small></p>
<p><small> Itabashi E, Osabe K, Fujimoto R, Kakizaki T (2017) Epigenetic regulation of agronomical traits in Brassicaceae. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2223-z</small></p>
<p><small> Kumar V et al (2017) Plant small RNAs: the essential epigenetic regulators of gene expression for salt-stress responses and tolerance. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2210-4</small></p>
<p><small> Law, J. A., and Jacobsen, S. E. (2010). Establishing, maintaining and modifying DNA methylation patterns in plants and animals. Nat. Rev. Genet. 11, 204–220. doi: 10.1038/nrg2719</small></p>
<p><small> Meyer, P. (2015). Epigenetic variation and environmental change. J. Exp. Bot. 66, 3541–3548. doi: 10.1093/jxb/eru502</small></p>
<p><small> Moazed, D. (2009) Small RNAs in transcriptional gene silencing and genome defence. Nature. doi: 10.1038/nature07756</small></p>
<p><small> Ong-Abdullah et al. (2015). Loss of Karma transposon methylation underlies the mantled somaclonal variant of oil palm. Nature 525, 533–537. doi: 10.1038/nature15365</small></p>
<p><small> Saéz-Laguna et al. (2014). Epigenetic variability in the genetically uniform forest tree species. PLoS One 9:e103145. doi: 10.1371/journal.pone.0103145</small></p>
<p><small> Sano, H. et al. (1990). A single treatment of rice seedlings with 5-azacytidine induces heritable dwarfism and undermethylation of genomic DNA. Mol. Gen. Genet. 220, 441–447. doi: 10.1007/BF00391751</small></p>
<p><small> Song, J et al (2012). Vernalization – A cold-induced epigenetic switch. J. Cell Sci. 125, 3723–3731. doi: 10.1242/jcs.084764</small></p>
<p><small> Steward, N et al. (2002). Periodic DNA methylation in maize nucleosomes and demethylation by environmental stress. J. Biol. Chem. 277, 37741–37746. doi: 10.1074/jbc.M204050200</small></p>
<p><small> Vaillant, I., and Paszkowski, J. (2007). Role of histone and DNA methylation in gene regulation. Curr. Opin. Plant Biol. 10, 528–533. doi: 10.1016/j.pbi.2007.06.008</small></p>
<p><small> Zhang, et al. (2006). Genome-wide high-resolution mapping and functional analysis of DNA methylation in Arabidopsis. Cell 126, 1189–1201. doi: 10.1016/j.cell.2006.08.003</small></p>
<p><small> Zhang et al. 2018 Understanding the evolutionary potential of epigenetic variation: a comparison of heritable phenotypic variation in epiRILs, RILs, and natural ecotypes of Arabidopsis thaliana. Heredity 121, 257–265 (2018) doi:10.1038/s41437-018-0095-9</small></p>
<p><small> Zheng X et al (2017) Histone acetylation is involved in GA-mediated 45S rDNA decondensation in maize aleurone layers. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2207-z</small></p>
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<p>Diagenode’s<span> </span><b>IPure</b><b><span> </span>kit<span> </span></b>is the only DNA purification kit using magnetic beads, that is specifically optimized for extracting DNA from<span> </span><b>ChIP</b><b>,<span> </span></b><b>MeDIP</b><span> </span>and<span> </span><b>CUT&Tag</b>. The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification. This simple and straightforward protocol delivers pure DNA ready for any downstream application (e.g. next generation sequencing). Comparing to phenol-chloroform extraction, the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples.</p>
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<h4>High DNA recovery after purification of ChIP samples using IPure technology</h4>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-chromatin-function.png" width="500" /></center>
<p></p>
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<li style="text-align: left;">Fast & easy to use</li>
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<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-A.png" alt="ChIP-seq figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-B.png" alt="ChIP-seq figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1.</strong> Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors (containing the IPure module for DNA purification) and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.</small></p>
<p></p>
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<p>Successful CUT&Tag results showing a low background with high region-specific enrichment has been generated using 50.000 of K562 cells, 1 µg of H3K4me3 or H3K27me3 antibody (Diagenode, C15410003 or C15410069, respectively) and proteinA-Tn5 (1:250) (Diagenode, C01070001). 1 µg of IgG (C15410206) was used as negative control. Samples were purified using the IPure kit v2 or phenol-chloroform purification. The below figures present the comparison of two purification methods.</p>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-fig2.png" style="display: block; margin-left: auto; margin-right: auto;" width="400" /></center><center>
<p style="text-align: center;"><small><strong>Figure 2.</strong> Heatmap 3kb upstream and downstream of the TSS for H3K4me3</small></p>
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<p><img src="https://www.diagenode.com/img/product/kits/ipure-fig3.png" style="display: block; margin-left: auto; margin-right: auto;" width="600" /></p>
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<center><small><strong>Figure 3.</strong> Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using Diagenode’s pA-Tn5 transposase (Cat. No. C01070002), H3K27me3 antibody (Cat. No. C15410069) and IPure kit v2 vs phenol chloroform purification (PC).</small></center>
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<center><img src="https://www.diagenode.com/img/product/kits/magmedip-seq-figure_multi3.jpg" alt="medip sequencing coverage" width="600" /></center><center></center><center>
<p></p>
<small><strong>Figure 4.</strong> Consistent coverage and methylation detection from different starting amounts of DNA with the Diagenode MagMeDIP-seq Package (including the Ipure kit for DNA purification). Samples containing decreasing starting amounts of DNA (from the top down: 1000 ng (red), 250 ng (blue), 100 ng (green)) originating from human blood were prepared, revealing a consistent coverage profile for the three different starting amounts, which enables reproducible methylation detection. The CpG islands (CGIs) (marked by yellow boxes in the bottom track) are predominantly unmethylated in the human genome, and as expected, we see a depletion of reads at and around CGIs.</small></center>
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<h5><strong>IPure after ChIP</strong></h5>
<p><strong>Step 1:</strong> Chromatin is decrosslinked and eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added.<br /> <strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet.<br /> <strong>Step 3:</strong> Proteins and remaining buffer are washed away.<br /> <strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after MeDIP</strong></h5>
<p><strong>Step 1:</strong> DNA is eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Remaining buffer are washed away.<br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after CUT&Tag</strong></h5>
<p><strong>Step 1:</strong> pA-Tn5 is inactivated and DNA released from the cells. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Proteins and remaining buffer are washed away. <br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).</p>
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<div class="small-6 columns"><center>Arabidopsis</center><center><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></center>
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<div class="small-6 columns"><center>Poplar</center><center><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></center>
<p><small><strong>Figure 3.</strong> ChIP-seq was performed on Populus trichocarpa stem differenciating xylem using the Premium H3K4me3 ChIP-seq grade antibody. Libraries were prepared with the <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the input and is considered as the background enrichment. The profile in red represents enrichments along a wide region of scaffold 18. Using the same scale, the peaks of the immunoprecipitated samples are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
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<div class="small-6 columns"><center>Tomato</center><center><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></center>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our Premium H3K4me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
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<div class="small-6 columns"><center>Maize</center><center><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></center>
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<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
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<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
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<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
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<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
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<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9-14ac-polyclonal-antibody-classic-sample-size-10-mg">H3K9/14ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
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<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27ac-polyclonal-antibody-premium-sample-size-10-ug">H3K27ac polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
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<div class="row">
<div class="small-6 columns"><center>Arabidopsis</center><center><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></center>
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<div class="small-6 columns"><center>Poplar</center><center><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></center>
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<div class="small-6 columns"><center>Tomato</center><center><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></center>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our Premium H3K4me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
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<div class="small-6 columns"><center>Maize</center><center><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></center>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our Premium H3K27me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
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<table style="width: 856px;">
<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
</td>
<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
</td>
<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
</td>
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<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27ac-polyclonal-antibody-premium-sample-size-10-ug">H3K27ac polyclonal antibody - Premium</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
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<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
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<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
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<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
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<tr>
<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
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<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the trimethylated lysine 4</strong> (<strong>H3K4me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4me3</strong><br />ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K4me3 (cat. No. C15410003) and optimized PCR primer pairs for qPCR. ChIP was performed with the iDeal ChIP-seq kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p></p>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2a-ChIP-seq.jpg" width="800" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2b-ChIP-seq.jpg" width="800" /></center><center>C.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2c-ChIP-seq.jpg" width="800" /></center><center>D.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2d-ChIP-seq.jpg" width="800" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K4me3</strong><br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of the Diagenode antibody against H3K4me3 (cat. No. C15410003) as described above. The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 600 kb region of the X-chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes, respectively (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</small></p>
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<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-cuttag-a.png" width="800" /></center></div>
<div class="small-12 columns"><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-cuttag-b.png" width="800" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K4me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 0.5 µg of the Diagenode antibody against H3K4me3 (cat. No. C15410003) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the ACTB gene on chromosome 7 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig3-ELISA.jpg" width="350" /></center><center></center><center></center><center></center><center></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody against H3K4me3 (cat. No. C15410003). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:11,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig4-DB.jpg" /></div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K4me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K4me3 (cat. No. C15410003), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 5A shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig5-WB.jpg" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K4me3</strong><br />Western blot was performed on whole cell extracts (40 µg, lane 1) from HeLa cells, and on 1 µg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K4me3 (cat. No. C15410003). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig6-if.jpg" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K4me3</strong><br />HeLa cells were stained with the Diagenode antibody against H3K4me3 (cat. No. C15410003) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. The cells were immunofluorescently labelled with the H3K4me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (middle), which specifically labels DNA. The right picture shows a merge of both stainings.</small></p>
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'info2' => '<p>Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called "histone code". Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Methylation of histone H3K4 is associated with activation of gene transcription.</p>
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'meta_title' => 'H3K4me3 Antibody - ChIP-seq Grade (C15410003) | Diagenode',
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'meta_description' => 'H3K4me3 (Histone H3 trimethylated at lysine 4) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array. Batch-specific data available on the website. Sample size available.',
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(int) 1 => array(
'id' => '2264',
'antibody_id' => '121',
'name' => 'H3K9me3 Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone<strong> H3 containing the trimethylated lysine 9</strong> (<strong>H3K9me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig1.png" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9me3</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K9me3 (cat. No. C15410193) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, 2, and 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for the heterochromatin marker Sat2 and for the ZNF510 gene, used as positive controls, and for the promoters of the active EIF4A2 and GAPDH genes, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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</div>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2a.png" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2b.png" width="700" /></center><center>C.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2c.png" width="700" /></center><center>D.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2d.png" width="700" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K9me3</strong><br />ChIP was performed with 0.5 µg of the Diagenode antibody against H3K9me3 (cat. No. C15410193) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2A shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. The arrows indicate two satellite repeat regions which exhibit a stronger signal. Figures 2B, 2C and 2D show the enrichment along the ZNF510 positive control target and at the H19 and KCNQ1 imprinted genes.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-CT-Fig3a.png" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-CT-Fig3b.png" width="700" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K9me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K9me3 (cat. No. C15410193) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in a genomic regions on chromosome 1 containing several ZNF repeat genes and in a genomic region surrounding the KCNQ1 imprinting control gene on chromosome 11 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-Elisa-Fig4.png" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human H3K9me3 (cat. No. C15410193) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:87,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-DB-Fig5.png" /></center></div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K9me3</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9me3 (cat. No. C15410193) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-WB-Fig6.png" /></center></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K9me3</strong><br />Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K9me3 (cat. No. C15410193). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-IF-Fig7.png" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K9me3</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9me3 (cat. No. C15410193) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me3 antibody (middle) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of both stainings is shown on the right.</small></p>
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'meta_description' => 'H3K9me3 (Histone H3 trimethylated at lysine 9) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array assay. Batch-specific data available on the website. Sample size available.',
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'name' => 'H3pan Antibody',
'description' => '<p><span>This antibody has been raised in rabbit against two KLH-conjugated synthetic peptides containing an unmodified sequence from the central part and from the C-terminus of <strong>histone H3</strong>, respectively.</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-chip.jpg" alt="H3pan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3pan</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3pan (Cat. No. C15310135) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 μl per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive MYOD1 and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-elisa.jpg" alt="H3pan Antibody ELISA validation" caption="false" width="288" height="217" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3pan (Cat. No. C15310135). The plates were coated with the peptides used for immunization. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be >1:1,000,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-wb.jpg" alt="H3pan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against H3pan</strong><br />Whole cell extracts from HeLa cells (25 μg) were analysed by Western blot using the Diagenode antibody against H3pan (Cat. No. C15310135) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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'meta_description' => 'H3pan (Histone H3) Polyclonal Antibody validated in ChIP-qPCR, WB and ELISA. Batch-specific data available on the website. Sample size available.',
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'name' => 'H3K27me3 Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the trimethylated lysine 27</strong> (<strong>H3K27me3</strong>), using a KLH-conjugated synthetic peptide.</p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig1.png" alt="H3K27me3 Antibody ChIP Grade" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2.png" alt="H3K27me3 Antibody for ChIP" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K27me3 (Cat. No. C15410195) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control.</small></p>
<p><small><strong>Figure 1A.</strong> Quantitative PCR was performed with primers specific for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive TSH2B and MYT1 genes, used as positive controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
<p><small><strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me3, H3K9me3 and H3K36me3 modifications and the unmodified H3K27 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H3K27me3 modification.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2a.png" alt="H3K27me3 Antibody ChIP-seq Grade" /></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-12 columns">
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2b.png" alt="H3K27me3 Antibody for ChIP-seq" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2c.png" alt="H3K27me3 Antibody for ChIP-seq assay" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2d.png" alt="H3K27me3 Antibody validated in ChIP-seq" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 µg of the Diagenode antibody against H3K27me3 (Cat. No. C15410195) as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment in genomic regions of chromosome 6 and 20, surrounding the TSH2B and MYT1 positive control genes (fig 2A and 2B, respectively), and in two genomic regions of chromosome 1 and X (figure 2C and D).</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-CUTTAG-Fig3A.png" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-CUTTAG-Fig3B.png" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K27me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K27me3 (cat. No. C15410195) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions on chromosome and 13 and 20 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-ELISA-Fig4.png" alt="H3K27me3 Antibody ELISA Validation " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me3 (Cat. No. C15410195). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:3,000.</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-DB-Fig5a.png" alt="H3K27me3 Antibody Dot Blot Validation " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K27me3</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3 (Cat. No. C15410195) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 5 shows a high specificity of the antibody for the modification of interest. Please note that the antibody also recognizes the modification if S28 is phosphorylated.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-WB-Fig6.png" alt="H3K27me3 Antibody validated in Western Blot" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K27me3 (cat. No. C15410195) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-IF-Fig7.png" alt="H3K27me3 Antibody validated for Immunofluorescence" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K27me3</strong><br />Human HeLa cells were stained with the Diagenode antibody against H3K27me3 (Cat. No. C15410195) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'slug' => 'h3k27me3-polyclonal-antibody-premium-50-mg-27-ml',
'meta_title' => 'H3K27me3 Antibody - ChIP-seq Grade (C15410195) | Diagenode',
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'meta_description' => 'H3K27me3 (Histone H3 trimethylated at lysine 27) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array assay. Batch-specific data available on the website. Sample size available.',
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'name' => 'Chromatin EasyShear Kit for Plant ',
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<p style="text-align: justify;"><span>Previous name of the kit: Chromatin Shearing Optimization Kit (Universal Plant ChIP-seq kit)<br /></span></p>
<p style="text-align: justify;"><span>The first critical step of a successful ChIP experiment is the best preparation of sheared chromatin. This <strong>Chromatin EasyShear Kit</strong> is designed to be used in conjunction with the <strong>Universal Plant ChIP-seq kit</strong> and contains the right level of <strong>detergent</strong> for extraction of highest quality plant chromatin for ChIP. In addition, the signature</span><span> crosslinking containers of this kit provide a simple and reliable method for fixation. The content of this kit is enough to perform 12 chromatin extractions.<br /></span></p>
<p style="text-align: justify;"><span>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
<p style="text-align: justify;"><span>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></span></p>',
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'slug' => 'chromatin-shearing-plant-chip-seq-kit',
'meta_title' => 'Chromatin Shearing Optimization Kit (Universal Plant ChIP-seq kit) | Diagenode',
'meta_keywords' => 'chromatin shearing, plant epigenetics, plant ChIP, plant ChIP-seq, Arabidopsis, maize, rice, tomato, poplar',
'meta_description' => 'Chromatin Shearing Optimization Kit designed to be used in conjunction with the Universal Plant ChIP-seq kit, providing high quality plant chromatin for ChIP',
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<p style="text-align: justify;">The <strong>Universal Plant ChIP-seq kit</strong> offers the convenience of extracting plant chromatin from a wide variety of plants including Arabidopsis, maize, rice, tomato and poplar. This complete kit has been specifically optimized for <strong>plant chromatin extraction</strong> and includes reagents for chromatin preparation, immunoprecipitation, plant-specific control primer pairs, control antibody, and DNA purification.</p>',
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<li><strong>Universal compatiblity</strong> with a wide variety of plant species</li>
<li>Optimized and <strong>complete kit</strong> for start-to-finish plant ChIP</li>
<li>Includes <strong>plant-specific control</strong> primers and control antibody<strong></strong></li>
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<h3>Successful ChIP-seq experiments for a variety of plants</h3>
<div class="row">
<div class="small-6 columns">
<h4 class="text-center">Arabidopsis</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG1"> <img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A-small.jpg" /> </a></p>
<div id="IMG1" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 1.</strong> ChIP-seq was performed on Arabidopsis thaliana (Col-0) seedlings using our <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng (green), 500 pg (orange) and 100 pg (red) IP'd DNA and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a public dataset (NCBI GEO Dataset GSM1193621) that we used as an external reference. Enrichments along a wide region of chromosome 5 are uniform regardless of the starting material amount for the preparation of the library.</small></p>
</div>
<div class="small-6 columns">
<h4 class="text-center">Poplar</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG2"><img src="https://www.diagenode.com/img/landing-pages/poplar-small.jpg" /> </a></p>
<div id="IMG2" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 3.</strong> ChIP-seq was performed on Populus trichocarpa stem differenciating xylem using the <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with the <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the input and is considered as the background enrichment. The profile in red represents enrichments along a wide region of scaffold 18. Using the same scale, the peaks of the immunoprecipitated samples are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<h4 class="text-center">Tomato</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG3"> <img src="https://www.diagenode.com/img/landing-pages/tomtato-small.jpg" /> </a></p>
<div id="IMG3" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
</div>
<div class="small-6 columns">
<h4 class="text-center">Maize</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG4"> <img src="https://www.diagenode.com/img/landing-pages/maize-small.jpg" /> </a></p>
<div id="IMG4" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our <a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">Premium H3K27me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<p><strong> </strong></p>
<table style="width: 856px;">
<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
</td>
<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
</td>
<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
</td>
</tr>
<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9-14ac-polyclonal-antibody-classic-sample-size-10-mg">H3K9/14ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27ac-polyclonal-antibody-premium-sample-size-10-ug">H3K27ac polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
</tbody>
</table>
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'meta_title' => 'Universal Plant ChIP-seq kit | Diagenode',
'meta_keywords' => 'plant epigenetics, plant ChIP, plant ChIP-seq, Arabidopsis, maize, rice, tomato, poplar',
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<p>Diagenode’s<span> </span><b>IPure</b><b><span> </span>kit<span> </span></b>is the only DNA purification kit using magnetic beads, that is specifically optimized for extracting DNA from<span> </span><b>ChIP</b><b>,<span> </span></b><b>MeDIP</b><span> </span>and<span> </span><b>CUT&Tag</b>. The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification. This simple and straightforward protocol delivers pure DNA ready for any downstream application (e.g. next generation sequencing). Comparing to phenol-chloroform extraction, the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples.</p>
<center>
<h4>High DNA recovery after purification of ChIP samples using IPure technology</h4>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-chromatin-function.png" width="500" /></center>
<p></p>
<p><small>ChIP assays were performed using different amounts of U2OS cells and the H3K9me3 antibody (Cat. No.<span> </span><span>C15410056</span>; 2 g/IP). <span>The purified DNA was eluted in 50 µl of water and quantified with a Nanodrop.</span></small></p>
<p></p>
<p><strong>Benefits of the IPure kit:</strong></p>
<ul>
<li style="text-align: left;">Provides pure DNA for any downstream application (e. g. Next generation sequencing)</li>
<li style="text-align: left;">Non-toxic</li>
<li style="text-align: left;">Fast & easy to use</li>
<li style="text-align: left;">Optimized for DNA purification after ChIP, MeDIP and CUT&Tag</li>
<li style="text-align: left;">Compatible with automation</li>
<li style="text-align: left;">Validated on the IP-Star Compact</li>
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'info1' => '<h2>IPure after ChIP</h2>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-A.png" alt="ChIP-seq figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-B.png" alt="ChIP-seq figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-C.png" alt="ChIP-seq figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small><strong>Figure 1.</strong> Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors (containing the IPure module for DNA purification) and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.</small></p>
<p></p>
<h2>IPure after CUT&Tag</h2>
<p>Successful CUT&Tag results showing a low background with high region-specific enrichment has been generated using 50.000 of K562 cells, 1 µg of H3K4me3 or H3K27me3 antibody (Diagenode, C15410003 or C15410069, respectively) and proteinA-Tn5 (1:250) (Diagenode, C01070001). 1 µg of IgG (C15410206) was used as negative control. Samples were purified using the IPure kit v2 or phenol-chloroform purification. The below figures present the comparison of two purification methods.</p>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-fig2.png" style="display: block; margin-left: auto; margin-right: auto;" width="400" /></center><center>
<p style="text-align: center;"><small><strong>Figure 2.</strong> Heatmap 3kb upstream and downstream of the TSS for H3K4me3</small></p>
</center>
<p></p>
<p><img src="https://www.diagenode.com/img/product/kits/ipure-fig3.png" style="display: block; margin-left: auto; margin-right: auto;" width="600" /></p>
<p></p>
<center><small><strong>Figure 3.</strong> Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using Diagenode’s pA-Tn5 transposase (Cat. No. C01070002), H3K27me3 antibody (Cat. No. C15410069) and IPure kit v2 vs phenol chloroform purification (PC).</small></center>
<p></p>
<p></p>
<h2>IPure after MeDIP</h2>
<center><img src="https://www.diagenode.com/img/product/kits/magmedip-seq-figure_multi3.jpg" alt="medip sequencing coverage" width="600" /></center><center></center><center>
<p></p>
<small><strong>Figure 4.</strong> Consistent coverage and methylation detection from different starting amounts of DNA with the Diagenode MagMeDIP-seq Package (including the Ipure kit for DNA purification). Samples containing decreasing starting amounts of DNA (from the top down: 1000 ng (red), 250 ng (blue), 100 ng (green)) originating from human blood were prepared, revealing a consistent coverage profile for the three different starting amounts, which enables reproducible methylation detection. The CpG islands (CGIs) (marked by yellow boxes in the bottom track) are predominantly unmethylated in the human genome, and as expected, we see a depletion of reads at and around CGIs.</small></center>
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<h3><strong>Workflow description</strong></h3>
<h5><strong>IPure after ChIP</strong></h5>
<p><strong>Step 1:</strong> Chromatin is decrosslinked and eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added.<br /> <strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet.<br /> <strong>Step 3:</strong> Proteins and remaining buffer are washed away.<br /> <strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after MeDIP</strong></h5>
<p><strong>Step 1:</strong> DNA is eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Remaining buffer are washed away.<br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after CUT&Tag</strong></h5>
<p><strong>Step 1:</strong> pA-Tn5 is inactivated and DNA released from the cells. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Proteins and remaining buffer are washed away. <br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).</p>
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'description' => '<div class="row">
<div class="large-12 columns">Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on a genome-wide scale. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</div>
<div class="large-12 columns"></div>
<h5 class="large-12 columns"><strong></strong></h5>
<h5 class="large-12 columns"><strong>The ChIP-seq workflow</strong></h5>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/chip-seq-diagram.png" /></div>
<div class="large-12 columns"><br />
<ol>
<li class="large-12 columns"><strong>Chromatin preparation: </strong>Crosslink chromatin-bound proteins (histones or transcription factors) to DNA followed by cell lysis.</li>
<li class="large-12 columns"><strong>Chromatin shearing:</strong> Fragment chromatin by sonication to desired fragment size (100-500 bp)</li>
<li class="large-12 columns"><strong>Chromatin IP</strong>: Capture protein-DNA complexes with <strong><a href="../categories/chip-seq-grade-antibodies">specific ChIP-seq grade antibodies</a></strong> against the histone or transcription factor of interest</li>
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<p class="text-justify">Chromatin Immunoprecipitation (ChIP) coupled with quantitative PCR can be used to investigate protein-DNA interaction at known genomic binding sites. if sites are not known, qPCR primers can also be designed against potential regulatory regions such as promoters. ChIP-qPCR is advantageous in studies that focus on specific genes and potential regulatory regions across differing experimental conditions as the cost of performing real-time PCR is minimal. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</p>
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<p><img src="https://www.diagenode.com/img/areas/plant.jpg" /></p>
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<h2>Epigenetic Regulation in Plants</h2>
<p>Plants utilize a number of gene regulation mechanisms to ensure proper development, function, growth, and survival under different environmental conditions. Plants depend on changes in gene expression to respond to environmental stimuli, in which the full repertoire of histone modifications, DNA methylation, and small ncRNAs play an important role in epigenetic regulation.</p>
<p>Studying the epigenetics of model plants such as Arabidopsis thaliana have allowed researchers to understand pathways that maintain chromatin modifications as well as the mapping of modifications such as DNA methylation on a genome-wide scale. Small RNAs have also been implicated in playing a role in the distribution of chromatin modifications, and RNA may also play a role in the complex epigenetic interactions that occur between homologous sequences (Moazed et al, 2009). In the future, by understanding epigenetic control, researchers can uncover the research necessary to improve plant growth, yields, and transformation efficiency especially in the face of climate change and other environmental factors.</p>
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<h3 style="font-weight: 100; margin-top: 0;">Chromatin</h3>
<p>Chromatin consists of nucleosomes formed by a complex of histone proteins and DNA, which allows the packaging of DNA into the nucleus. The less condensed euchromatin represents transcriptionally active regions, while heterochromatin is usually inactive (Vaillant and Paszkowski, 2007). Chromatin state is known to be influenced by both DNA methylation and histone modifications which in turn impact gene expression and the structure of chromosomes. In a recent study, the role of chromatin modifications during plant reproduction elucidated 3-dimensional chromosome reorganization mediated by histones and DNA methylation (Dukowic-Schulze et al. 2017). In addition, gibberellins have been shown in increasing the level of histone acetylation, which affects regions of chromatin involved in maize seed germination (Zheng et al. 2017). Another study reports a novel function of a tomato histone deacetylase gene in the regulation of fruit ripening (Guo et al. 2017).</p>
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<p>In addition, multigene families encode transcription factors, with members found throughout the genome or clustered on the same chromosome. Numerous DNA binding proteins that interact with plant promoters have been identified -- some are similar to well-characterized transcription factors in animals or yeast, while others are unique to plants. For example, diverse members of the subfamily X of the plant-specific ethylene response factor (ERF) transcription factors coordinate stress signaling with wound repair activation. Tissue repair is also enhanced through a protein complex of ERF and GRAS TFs (Heyman et. al,.2018). A compilation of known plant transcription factors can be found in the plant transcription factor database at http://plntfdb.bio.uni-potsdam.de/v3.0/.</p>
</div>
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<p><img src="https://www.diagenode.com/img/areas/rna-strand.jpg" /></p>
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<div class="small-12 medium-9 large-9 columns">
<h3 style="font-weight: 100; margin-top: 0;">RNA</h3>
<p>Recent research shows that a number of classes of small RNAs are key epigenetic regulators. In many cases, small RNAs have been implicated in DNA methylation and chromatin modification (Meyer, 2015). In addition, the role of small RNAs has been implicated in plant stress tolerance (Kumar et al., 2017). López-Galiano et al also provided insight into a coordinated function of a miRNA gene and histone modifications in regulating the expression of a WRKY transcription factor in response to stress.</p>
<p>RNA interference (RNAi) is another epigenetic mechanism that leads to small RNA generation, which mediates gene silencing at the post-transcriptional level. RNAi technology has immense potential for plant disease resistance.</p>
</div>
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<p><img src="https://www.diagenode.com/img/areas/dna-methylation.jpg" /></p>
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<div class="small-12 medium-9 large-9 columns">
<h3 style="font-weight: 100; margin-top: 0;">DNA methylation</h3>
<p>Plants, unlike animals, have three sites that can be methylated G, CHG (H can be A, C, T), and CHH (Law and Jacobsen, 2010). DNA methylation has attracted particular interest. In Arabidopsis, one-third of methylated genes occur in transcribed regions, and 5% of genes are methylated in promoter regions, suggesting that many of these are epigenetically regulated. (Zhang et al., 2006).</p>
<p>There are thousands of differentially methylated regions (DMRs) that influence phenotype by influencing gene expression. The analysis of epigenetic recombinant inbred line (epiRIL) plants from Arabidopsis points to the evidence of the influence of DMRs. An epiRIL results from crossing two genetically identical plants with differing DNA methylation levels (with one parent as a homozygous mutant for an essential DNA methylation maintenance gene). The offspring of these plants have similar genomes that vary only in methylation levels. Many traits have been studied using epiRILs -- flowering time, plant height, and response to abiotic stress, some of which have now been mapped to DMRs (Zhang et al. 2018)</p>
<p>Regulation by DNA methylation has been shown to be important in many aspects of plant development and response such as vernalization, hybrid vigor, and self-incompatibility (Itabashi et al. 2017). For example, vernalization treatments have shown reduced DNA methylation and subsequent initiation of flowering (Burn et al., 1993). Stress can also influence DNA methylation in plants as a response to environmental stimuli. (Steward et al., 2002; Song et al., 2012). A high degree of DNA methylation has also suggested the role in the improvement of plant fitness under different environmental conditions (Saéz-Laguna et al., 2014). In addition, methylation can affect normal fruit and hypomethylation predicts homeotic transformation and loss of fruit yield (Ong-Abdullah et al., 2015)</p>
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<p>DNA demethylation has also been implied in various aspects of plant development including pollen tube formation, embryogenesis, fruit ripening, stomatal development, and nodule formation ( Li et al. 2017). Demethylation of rice genomic DNA caused an altered pattern of gene expression, inducing dwarf plants (Sano et al., 1990).</p>
<p>Epigenetic modifications contribute to the stability and survival of the plants and their ability to adapt in different environmental conditions.</p>
</div>
</div>
<h3>Diagenode products for your epigenomics research in plants</h3>
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<h3 class="text-center"><a href="https://www.diagenode.com/en/categories/chromatin-function">Chromatin analysis</a></h3>
<center><a href="https://www.diagenode.com/en/categories/chromatin-function"><img src="https://www.diagenode.com/img/cancer/chromatin-icon.png" /></a></center>
<p class="text-left">Understand the role of chromatin in plant function and development</p>
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<li><a href="https://www.diagenode.com/en/categories/chromatin-function">Learn about our chromatin analysis products</a></li>
<li><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns"> Learn about the Universal Plant ChIP Kit</a></li>
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<div class="small-12 medium-4 large-4 columns text-left">
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<h3 class="text-center"><a href="https://www.diagenode.com/en/categories/dna-methylation" style="color: #30415c;">DNA methylation</a></h3>
<center><a href="https://www.diagenode.com/en/categories/dna-methylation"><img src="https://www.diagenode.com/img/cancer/dna-icon.png" /></a></center>
<p class="text-left">DNA methylation and demethylation and the effects on plant response and function</p>
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<li><a href="https://www.diagenode.com/en/categories/dna-methylation">Discover DNA methylation analysis solutions at any resolution</a></li>
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<div class="panel" style="border-color: #474546; height: 275px;">
<h3 class="text-center"><span class="darkgrey">Non-coding RNAs</span></h3>
<center><img src="https://www.diagenode.com/img/cancer/non-coding-icon.png" /></center>
<p class="text-left">Discover noncoding RNAs in the regulation of gene expression in plants</p>
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<li><a href="https://www.diagenode.com/en/categories/Library-preparation-for-RNA-seq">Library prep for RNA-seq studies for ncRNAs</a></li>
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<h3>References</h3>
<p><small> Burn, J. et al (1993). DNA methylation, vernalization, and the initiation of flowering. Proc. Natl. Acad. Sci. U.S.A. 90, 287–291. doi: 10.1006/scdb.1996.0055 </small></p>
<p><small> Dukowic-Schulze S, Liu C, Chen C (2017) Not just gene expression: 3D implications of chromatin modifications during sexual plant reproduction. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2222-0</small></p>
<p><small> Guo J et al (2017) A histone deacetylase gene, SlHDA3, acts as a negative regulator of fruit ripening and carotenoid accumulation. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2211-3</small></p>
<p><small> Heyman J, et.al (2018) Journal of Cell Science Emerging role of the plant ERF transcription factors in coordinating wound defense responses and repair doi: 10.1242/jcs.208215</small></p>
<p><small> Itabashi E, Osabe K, Fujimoto R, Kakizaki T (2017) Epigenetic regulation of agronomical traits in Brassicaceae. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2223-z</small></p>
<p><small> Kumar V et al (2017) Plant small RNAs: the essential epigenetic regulators of gene expression for salt-stress responses and tolerance. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2210-4</small></p>
<p><small> Law, J. A., and Jacobsen, S. E. (2010). Establishing, maintaining and modifying DNA methylation patterns in plants and animals. Nat. Rev. Genet. 11, 204–220. doi: 10.1038/nrg2719</small></p>
<p><small> Meyer, P. (2015). Epigenetic variation and environmental change. J. Exp. Bot. 66, 3541–3548. doi: 10.1093/jxb/eru502</small></p>
<p><small> Moazed, D. (2009) Small RNAs in transcriptional gene silencing and genome defence. Nature. doi: 10.1038/nature07756</small></p>
<p><small> Ong-Abdullah et al. (2015). Loss of Karma transposon methylation underlies the mantled somaclonal variant of oil palm. Nature 525, 533–537. doi: 10.1038/nature15365</small></p>
<p><small> Saéz-Laguna et al. (2014). Epigenetic variability in the genetically uniform forest tree species. PLoS One 9:e103145. doi: 10.1371/journal.pone.0103145</small></p>
<p><small> Sano, H. et al. (1990). A single treatment of rice seedlings with 5-azacytidine induces heritable dwarfism and undermethylation of genomic DNA. Mol. Gen. Genet. 220, 441–447. doi: 10.1007/BF00391751</small></p>
<p><small> Song, J et al (2012). Vernalization – A cold-induced epigenetic switch. J. Cell Sci. 125, 3723–3731. doi: 10.1242/jcs.084764</small></p>
<p><small> Steward, N et al. (2002). Periodic DNA methylation in maize nucleosomes and demethylation by environmental stress. J. Biol. Chem. 277, 37741–37746. doi: 10.1074/jbc.M204050200</small></p>
<p><small> Vaillant, I., and Paszkowski, J. (2007). Role of histone and DNA methylation in gene regulation. Curr. Opin. Plant Biol. 10, 528–533. doi: 10.1016/j.pbi.2007.06.008</small></p>
<p><small> Zhang, et al. (2006). Genome-wide high-resolution mapping and functional analysis of DNA methylation in Arabidopsis. Cell 126, 1189–1201. doi: 10.1016/j.cell.2006.08.003</small></p>
<p><small> Zhang et al. 2018 Understanding the evolutionary potential of epigenetic variation: a comparison of heritable phenotypic variation in epiRILs, RILs, and natural ecotypes of Arabidopsis thaliana. Heredity 121, 257–265 (2018) doi:10.1038/s41437-018-0095-9</small></p>
<p><small> Zheng X et al (2017) Histone acetylation is involved in GA-mediated 45S rDNA decondensation in maize aleurone layers. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2207-z</small></p>
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<p>Diagenode’s<span> </span><b>IPure</b><b><span> </span>kit<span> </span></b>is the only DNA purification kit using magnetic beads, that is specifically optimized for extracting DNA from<span> </span><b>ChIP</b><b>,<span> </span></b><b>MeDIP</b><span> </span>and<span> </span><b>CUT&Tag</b>. The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification. This simple and straightforward protocol delivers pure DNA ready for any downstream application (e.g. next generation sequencing). Comparing to phenol-chloroform extraction, the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples.</p>
<center>
<h4>High DNA recovery after purification of ChIP samples using IPure technology</h4>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-chromatin-function.png" width="500" /></center>
<p></p>
<p><small>ChIP assays were performed using different amounts of U2OS cells and the H3K9me3 antibody (Cat. No.<span> </span><span>C15410056</span>; 2 g/IP). <span>The purified DNA was eluted in 50 µl of water and quantified with a Nanodrop.</span></small></p>
<p></p>
<p><strong>Benefits of the IPure kit:</strong></p>
<ul>
<li style="text-align: left;">Provides pure DNA for any downstream application (e. g. Next generation sequencing)</li>
<li style="text-align: left;">Non-toxic</li>
<li style="text-align: left;">Fast & easy to use</li>
<li style="text-align: left;">Optimized for DNA purification after ChIP, MeDIP and CUT&Tag</li>
<li style="text-align: left;">Compatible with automation</li>
<li style="text-align: left;">Validated on the IP-Star Compact</li>
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'info1' => '<h2>IPure after ChIP</h2>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-A.png" alt="ChIP-seq figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-B.png" alt="ChIP-seq figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-C.png" alt="ChIP-seq figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small><strong>Figure 1.</strong> Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors (containing the IPure module for DNA purification) and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.</small></p>
<p></p>
<h2>IPure after CUT&Tag</h2>
<p>Successful CUT&Tag results showing a low background with high region-specific enrichment has been generated using 50.000 of K562 cells, 1 µg of H3K4me3 or H3K27me3 antibody (Diagenode, C15410003 or C15410069, respectively) and proteinA-Tn5 (1:250) (Diagenode, C01070001). 1 µg of IgG (C15410206) was used as negative control. Samples were purified using the IPure kit v2 or phenol-chloroform purification. The below figures present the comparison of two purification methods.</p>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-fig2.png" style="display: block; margin-left: auto; margin-right: auto;" width="400" /></center><center>
<p style="text-align: center;"><small><strong>Figure 2.</strong> Heatmap 3kb upstream and downstream of the TSS for H3K4me3</small></p>
</center>
<p></p>
<p><img src="https://www.diagenode.com/img/product/kits/ipure-fig3.png" style="display: block; margin-left: auto; margin-right: auto;" width="600" /></p>
<p></p>
<center><small><strong>Figure 3.</strong> Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using Diagenode’s pA-Tn5 transposase (Cat. No. C01070002), H3K27me3 antibody (Cat. No. C15410069) and IPure kit v2 vs phenol chloroform purification (PC).</small></center>
<p></p>
<p></p>
<h2>IPure after MeDIP</h2>
<center><img src="https://www.diagenode.com/img/product/kits/magmedip-seq-figure_multi3.jpg" alt="medip sequencing coverage" width="600" /></center><center></center><center>
<p></p>
<small><strong>Figure 4.</strong> Consistent coverage and methylation detection from different starting amounts of DNA with the Diagenode MagMeDIP-seq Package (including the Ipure kit for DNA purification). Samples containing decreasing starting amounts of DNA (from the top down: 1000 ng (red), 250 ng (blue), 100 ng (green)) originating from human blood were prepared, revealing a consistent coverage profile for the three different starting amounts, which enables reproducible methylation detection. The CpG islands (CGIs) (marked by yellow boxes in the bottom track) are predominantly unmethylated in the human genome, and as expected, we see a depletion of reads at and around CGIs.</small></center>
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'label2' => 'iPure Workflow',
'info2' => '<h2 style="text-align: center;">Kit Method Overview & Time table</h2>
<p><img src="https://www.diagenode.com/img/product/kits/workflow-ipure-cuttag.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h3><strong>Workflow description</strong></h3>
<h5><strong>IPure after ChIP</strong></h5>
<p><strong>Step 1:</strong> Chromatin is decrosslinked and eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added.<br /> <strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet.<br /> <strong>Step 3:</strong> Proteins and remaining buffer are washed away.<br /> <strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after MeDIP</strong></h5>
<p><strong>Step 1:</strong> DNA is eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Remaining buffer are washed away.<br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after CUT&Tag</strong></h5>
<p><strong>Step 1:</strong> pA-Tn5 is inactivated and DNA released from the cells. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Proteins and remaining buffer are washed away. <br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).</p>
<p></p>
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<div class="small-6 columns"><center>Poplar</center><center><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></center>
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<div class="small-6 columns"><center>Tomato</center><center><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></center>
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<div class="small-6 columns"><center>Maize</center><center><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></center>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
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<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
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<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
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<div class="small-6 columns"><center>Arabidopsis</center><center><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></center>
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<div class="small-6 columns"><center>Poplar</center><center><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></center>
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<div class="small-6 columns"><center>Tomato</center><center><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></center>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our Premium H3K4me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
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<div class="small-6 columns"><center>Maize</center><center><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></center>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our Premium H3K27me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
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<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
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<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
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<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
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<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
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<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
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<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
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<tr>
<td style="width: 224px;"></td>
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<td style="width: 357px;"></td>
</tr>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the trimethylated lysine 4</strong> (<strong>H3K4me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4me3</strong><br />ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K4me3 (cat. No. C15410003) and optimized PCR primer pairs for qPCR. ChIP was performed with the iDeal ChIP-seq kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p></p>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2a-ChIP-seq.jpg" width="800" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2b-ChIP-seq.jpg" width="800" /></center><center>C.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2c-ChIP-seq.jpg" width="800" /></center><center>D.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2d-ChIP-seq.jpg" width="800" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K4me3</strong><br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of the Diagenode antibody against H3K4me3 (cat. No. C15410003) as described above. The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 600 kb region of the X-chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes, respectively (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-cuttag-a.png" width="800" /></center></div>
<div class="small-12 columns"><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-cuttag-b.png" width="800" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K4me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 0.5 µg of the Diagenode antibody against H3K4me3 (cat. No. C15410003) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the ACTB gene on chromosome 7 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig3-ELISA.jpg" width="350" /></center><center></center><center></center><center></center><center></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody against H3K4me3 (cat. No. C15410003). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:11,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig4-DB.jpg" /></div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K4me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K4me3 (cat. No. C15410003), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 5A shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig5-WB.jpg" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K4me3</strong><br />Western blot was performed on whole cell extracts (40 µg, lane 1) from HeLa cells, and on 1 µg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K4me3 (cat. No. C15410003). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig6-if.jpg" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K4me3</strong><br />HeLa cells were stained with the Diagenode antibody against H3K4me3 (cat. No. C15410003) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. The cells were immunofluorescently labelled with the H3K4me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (middle), which specifically labels DNA. The right picture shows a merge of both stainings.</small></p>
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'info2' => '<p>Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called "histone code". Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Methylation of histone H3K4 is associated with activation of gene transcription.</p>
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'meta_description' => 'H3K4me3 (Histone H3 trimethylated at lysine 4) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array. Batch-specific data available on the website. Sample size available.',
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'id' => '2264',
'antibody_id' => '121',
'name' => 'H3K9me3 Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone<strong> H3 containing the trimethylated lysine 9</strong> (<strong>H3K9me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig1.png" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9me3</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K9me3 (cat. No. C15410193) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, 2, and 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for the heterochromatin marker Sat2 and for the ZNF510 gene, used as positive controls, and for the promoters of the active EIF4A2 and GAPDH genes, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2a.png" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2b.png" width="700" /></center><center>C.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2c.png" width="700" /></center><center>D.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-ChIP-Fig2d.png" width="700" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K9me3</strong><br />ChIP was performed with 0.5 µg of the Diagenode antibody against H3K9me3 (cat. No. C15410193) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2A shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. The arrows indicate two satellite repeat regions which exhibit a stronger signal. Figures 2B, 2C and 2D show the enrichment along the ZNF510 positive control target and at the H19 and KCNQ1 imprinted genes.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-CT-Fig3a.png" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-CT-Fig3b.png" width="700" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K9me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K9me3 (cat. No. C15410193) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in a genomic regions on chromosome 1 containing several ZNF repeat genes and in a genomic region surrounding the KCNQ1 imprinting control gene on chromosome 11 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-Elisa-Fig4.png" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human H3K9me3 (cat. No. C15410193) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:87,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-DB-Fig5.png" /></center></div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K9me3</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9me3 (cat. No. C15410193) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
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<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-WB-Fig6.png" /></center></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K9me3</strong><br />Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K9me3 (cat. No. C15410193). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-IF-Fig7.png" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K9me3</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9me3 (cat. No. C15410193) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me3 antibody (middle) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of both stainings is shown on the right.</small></p>
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'meta_description' => 'H3K9me3 (Histone H3 trimethylated at lysine 9) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array assay. Batch-specific data available on the website. Sample size available.',
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'name' => 'H3pan Antibody',
'description' => '<p><span>This antibody has been raised in rabbit against two KLH-conjugated synthetic peptides containing an unmodified sequence from the central part and from the C-terminus of <strong>histone H3</strong>, respectively.</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-chip.jpg" alt="H3pan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3pan</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3pan (Cat. No. C15310135) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 μl per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive MYOD1 and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-elisa.jpg" alt="H3pan Antibody ELISA validation" caption="false" width="288" height="217" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3pan (Cat. No. C15310135). The plates were coated with the peptides used for immunization. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be >1:1,000,000.</small></p>
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</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-wb.jpg" alt="H3pan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against H3pan</strong><br />Whole cell extracts from HeLa cells (25 μg) were analysed by Western blot using the Diagenode antibody against H3pan (Cat. No. C15310135) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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'meta_description' => 'H3pan (Histone H3) Polyclonal Antibody validated in ChIP-qPCR, WB and ELISA. Batch-specific data available on the website. Sample size available.',
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'id' => '2268',
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'name' => 'H3K27me3 Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the trimethylated lysine 27</strong> (<strong>H3K27me3</strong>), using a KLH-conjugated synthetic peptide.</p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig1.png" alt="H3K27me3 Antibody ChIP Grade" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2.png" alt="H3K27me3 Antibody for ChIP" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K27me3 (Cat. No. C15410195) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control.</small></p>
<p><small><strong>Figure 1A.</strong> Quantitative PCR was performed with primers specific for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive TSH2B and MYT1 genes, used as positive controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
<p><small><strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me3, H3K9me3 and H3K36me3 modifications and the unmodified H3K27 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H3K27me3 modification.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2a.png" alt="H3K27me3 Antibody ChIP-seq Grade" /></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-12 columns">
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2b.png" alt="H3K27me3 Antibody for ChIP-seq" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2c.png" alt="H3K27me3 Antibody for ChIP-seq assay" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2d.png" alt="H3K27me3 Antibody validated in ChIP-seq" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 µg of the Diagenode antibody against H3K27me3 (Cat. No. C15410195) as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment in genomic regions of chromosome 6 and 20, surrounding the TSH2B and MYT1 positive control genes (fig 2A and 2B, respectively), and in two genomic regions of chromosome 1 and X (figure 2C and D).</small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-CUTTAG-Fig3A.png" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-CUTTAG-Fig3B.png" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K27me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K27me3 (cat. No. C15410195) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions on chromosome and 13 and 20 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-ELISA-Fig4.png" alt="H3K27me3 Antibody ELISA Validation " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me3 (Cat. No. C15410195). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:3,000.</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-DB-Fig5a.png" alt="H3K27me3 Antibody Dot Blot Validation " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K27me3</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3 (Cat. No. C15410195) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 5 shows a high specificity of the antibody for the modification of interest. Please note that the antibody also recognizes the modification if S28 is phosphorylated.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-WB-Fig6.png" alt="H3K27me3 Antibody validated in Western Blot" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K27me3 (cat. No. C15410195) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-IF-Fig7.png" alt="H3K27me3 Antibody validated for Immunofluorescence" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K27me3</strong><br />Human HeLa cells were stained with the Diagenode antibody against H3K27me3 (Cat. No. C15410195) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'meta_title' => 'H3K27me3 Antibody - ChIP-seq Grade (C15410195) | Diagenode',
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'meta_description' => 'H3K27me3 (Histone H3 trimethylated at lysine 27) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, CUT&Tag, ELISA, DB, WB and IF. Specificity confirmed by Peptide array assay. Batch-specific data available on the website. Sample size available.',
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<p style="text-align: justify;"><span>Previous name of the kit: Chromatin Shearing Optimization Kit (Universal Plant ChIP-seq kit)<br /></span></p>
<p style="text-align: justify;"><span>The first critical step of a successful ChIP experiment is the best preparation of sheared chromatin. This <strong>Chromatin EasyShear Kit</strong> is designed to be used in conjunction with the <strong>Universal Plant ChIP-seq kit</strong> and contains the right level of <strong>detergent</strong> for extraction of highest quality plant chromatin for ChIP. In addition, the signature</span><span> crosslinking containers of this kit provide a simple and reliable method for fixation. The content of this kit is enough to perform 12 chromatin extractions.<br /></span></p>
<p style="text-align: justify;"><span>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
<p style="text-align: justify;"><span>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></span></p>',
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<p style="text-align: justify;">The <strong>Universal Plant ChIP-seq kit</strong> offers the convenience of extracting plant chromatin from a wide variety of plants including Arabidopsis, maize, rice, tomato and poplar. This complete kit has been specifically optimized for <strong>plant chromatin extraction</strong> and includes reagents for chromatin preparation, immunoprecipitation, plant-specific control primer pairs, control antibody, and DNA purification.</p>',
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<li><strong>Universal compatiblity</strong> with a wide variety of plant species</li>
<li>Optimized and <strong>complete kit</strong> for start-to-finish plant ChIP</li>
<li>Includes <strong>plant-specific control</strong> primers and control antibody<strong></strong></li>
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<h3>Successful ChIP-seq experiments for a variety of plants</h3>
<div class="row">
<div class="small-6 columns">
<h4 class="text-center">Arabidopsis</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG1"> <img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A-small.jpg" /> </a></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 1.</strong> ChIP-seq was performed on Arabidopsis thaliana (Col-0) seedlings using our <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng (green), 500 pg (orange) and 100 pg (red) IP'd DNA and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a public dataset (NCBI GEO Dataset GSM1193621) that we used as an external reference. Enrichments along a wide region of chromosome 5 are uniform regardless of the starting material amount for the preparation of the library.</small></p>
</div>
<div class="small-6 columns">
<h4 class="text-center">Poplar</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG2"><img src="https://www.diagenode.com/img/landing-pages/poplar-small.jpg" /> </a></p>
<div id="IMG2" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 3.</strong> ChIP-seq was performed on Populus trichocarpa stem differenciating xylem using the <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with the <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the input and is considered as the background enrichment. The profile in red represents enrichments along a wide region of scaffold 18. Using the same scale, the peaks of the immunoprecipitated samples are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<h4 class="text-center">Tomato</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG3"> <img src="https://www.diagenode.com/img/landing-pages/tomtato-small.jpg" /> </a></p>
<div id="IMG3" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
</div>
<div class="small-6 columns">
<h4 class="text-center">Maize</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG4"> <img src="https://www.diagenode.com/img/landing-pages/maize-small.jpg" /> </a></p>
<div id="IMG4" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our <a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">Premium H3K27me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<p><strong> </strong></p>
<table style="width: 856px;">
<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
</td>
<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
</td>
<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
</td>
</tr>
<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9-14ac-polyclonal-antibody-classic-sample-size-10-mg">H3K9/14ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27ac-polyclonal-antibody-premium-sample-size-10-ug">H3K27ac polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
</tbody>
</table>
<p><strong></strong></p>
<p><strong></strong></p>',
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'meta_title' => 'Universal Plant ChIP-seq kit | Diagenode',
'meta_keywords' => 'plant epigenetics, plant ChIP, plant ChIP-seq, Arabidopsis, maize, rice, tomato, poplar',
'meta_description' => 'Optimized extraction of plant chromatin from Arabidopsis,maize,rice,tomato,poplar.Complete ChIP kit including plant-specific control primer pairs and antibody',
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<p>Diagenode’s<span> </span><b>IPure</b><b><span> </span>kit<span> </span></b>is the only DNA purification kit using magnetic beads, that is specifically optimized for extracting DNA from<span> </span><b>ChIP</b><b>,<span> </span></b><b>MeDIP</b><span> </span>and<span> </span><b>CUT&Tag</b>. The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification. This simple and straightforward protocol delivers pure DNA ready for any downstream application (e.g. next generation sequencing). Comparing to phenol-chloroform extraction, the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples.</p>
<center>
<h4>High DNA recovery after purification of ChIP samples using IPure technology</h4>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-chromatin-function.png" width="500" /></center>
<p></p>
<p><small>ChIP assays were performed using different amounts of U2OS cells and the H3K9me3 antibody (Cat. No.<span> </span><span>C15410056</span>; 2 g/IP). <span>The purified DNA was eluted in 50 µl of water and quantified with a Nanodrop.</span></small></p>
<p></p>
<p><strong>Benefits of the IPure kit:</strong></p>
<ul>
<li style="text-align: left;">Provides pure DNA for any downstream application (e. g. Next generation sequencing)</li>
<li style="text-align: left;">Non-toxic</li>
<li style="text-align: left;">Fast & easy to use</li>
<li style="text-align: left;">Optimized for DNA purification after ChIP, MeDIP and CUT&Tag</li>
<li style="text-align: left;">Compatible with automation</li>
<li style="text-align: left;">Validated on the IP-Star Compact</li>
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'info1' => '<h2>IPure after ChIP</h2>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-A.png" alt="ChIP-seq figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-B.png" alt="ChIP-seq figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-C.png" alt="ChIP-seq figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small><strong>Figure 1.</strong> Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors (containing the IPure module for DNA purification) and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.</small></p>
<p></p>
<h2>IPure after CUT&Tag</h2>
<p>Successful CUT&Tag results showing a low background with high region-specific enrichment has been generated using 50.000 of K562 cells, 1 µg of H3K4me3 or H3K27me3 antibody (Diagenode, C15410003 or C15410069, respectively) and proteinA-Tn5 (1:250) (Diagenode, C01070001). 1 µg of IgG (C15410206) was used as negative control. Samples were purified using the IPure kit v2 or phenol-chloroform purification. The below figures present the comparison of two purification methods.</p>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-fig2.png" style="display: block; margin-left: auto; margin-right: auto;" width="400" /></center><center>
<p style="text-align: center;"><small><strong>Figure 2.</strong> Heatmap 3kb upstream and downstream of the TSS for H3K4me3</small></p>
</center>
<p></p>
<p><img src="https://www.diagenode.com/img/product/kits/ipure-fig3.png" style="display: block; margin-left: auto; margin-right: auto;" width="600" /></p>
<p></p>
<center><small><strong>Figure 3.</strong> Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using Diagenode’s pA-Tn5 transposase (Cat. No. C01070002), H3K27me3 antibody (Cat. No. C15410069) and IPure kit v2 vs phenol chloroform purification (PC).</small></center>
<p></p>
<p></p>
<h2>IPure after MeDIP</h2>
<center><img src="https://www.diagenode.com/img/product/kits/magmedip-seq-figure_multi3.jpg" alt="medip sequencing coverage" width="600" /></center><center></center><center>
<p></p>
<small><strong>Figure 4.</strong> Consistent coverage and methylation detection from different starting amounts of DNA with the Diagenode MagMeDIP-seq Package (including the Ipure kit for DNA purification). Samples containing decreasing starting amounts of DNA (from the top down: 1000 ng (red), 250 ng (blue), 100 ng (green)) originating from human blood were prepared, revealing a consistent coverage profile for the three different starting amounts, which enables reproducible methylation detection. The CpG islands (CGIs) (marked by yellow boxes in the bottom track) are predominantly unmethylated in the human genome, and as expected, we see a depletion of reads at and around CGIs.</small></center>
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<h3><strong>Workflow description</strong></h3>
<h5><strong>IPure after ChIP</strong></h5>
<p><strong>Step 1:</strong> Chromatin is decrosslinked and eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added.<br /> <strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet.<br /> <strong>Step 3:</strong> Proteins and remaining buffer are washed away.<br /> <strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after MeDIP</strong></h5>
<p><strong>Step 1:</strong> DNA is eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Remaining buffer are washed away.<br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after CUT&Tag</strong></h5>
<p><strong>Step 1:</strong> pA-Tn5 is inactivated and DNA released from the cells. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Proteins and remaining buffer are washed away. <br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).</p>
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<div class="large-12 columns">Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on a genome-wide scale. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</div>
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<h5 class="large-12 columns"><strong></strong></h5>
<h5 class="large-12 columns"><strong>The ChIP-seq workflow</strong></h5>
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<li class="large-12 columns"><strong>Chromatin preparation: </strong>Crosslink chromatin-bound proteins (histones or transcription factors) to DNA followed by cell lysis.</li>
<li class="large-12 columns"><strong>Chromatin shearing:</strong> Fragment chromatin by sonication to desired fragment size (100-500 bp)</li>
<li class="large-12 columns"><strong>Chromatin IP</strong>: Capture protein-DNA complexes with <strong><a href="../categories/chip-seq-grade-antibodies">specific ChIP-seq grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: Reverse cross-links, elute, and purify </li>
<li class="large-12 columns"><strong>NGS Library Preparation</strong>: Ligate adapters and amplify IP'd material</li>
<li class="large-12 columns"><strong>Bioinformatic analysis</strong>: Perform r<span style="font-weight: 400;">ead filtering and trimming</span>, r<span style="font-weight: 400;">ead specific alignment, enrichment specific peak calling, QC metrics, multi-sample cross-comparison etc. </span></li>
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<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
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<p class="text-justify">Chromatin Immunoprecipitation (ChIP) coupled with quantitative PCR can be used to investigate protein-DNA interaction at known genomic binding sites. if sites are not known, qPCR primers can also be designed against potential regulatory regions such as promoters. ChIP-qPCR is advantageous in studies that focus on specific genes and potential regulatory regions across differing experimental conditions as the cost of performing real-time PCR is minimal. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</p>
<p class="text-justify"><strong>The ChIP-qPCR workflow</strong></p>
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<li class="large-12 columns"><strong>Chromatin IP</strong>: protein-DNA complexe capture using<strong> <a href="https://www.diagenode.com/en/categories/chip-grade-antibodies">specific ChIP-grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: chromatin reverse cross-linking and elution followed by purification<strong> </strong></li>
<li class="large-12 columns"><strong>qPCR and analysis</strong>: using previously designed primers to amplify IP'd material at specific loci</li>
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<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
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<p><img src="https://www.diagenode.com/img/areas/plant.jpg" /></p>
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<h2>Epigenetic Regulation in Plants</h2>
<p>Plants utilize a number of gene regulation mechanisms to ensure proper development, function, growth, and survival under different environmental conditions. Plants depend on changes in gene expression to respond to environmental stimuli, in which the full repertoire of histone modifications, DNA methylation, and small ncRNAs play an important role in epigenetic regulation.</p>
<p>Studying the epigenetics of model plants such as Arabidopsis thaliana have allowed researchers to understand pathways that maintain chromatin modifications as well as the mapping of modifications such as DNA methylation on a genome-wide scale. Small RNAs have also been implicated in playing a role in the distribution of chromatin modifications, and RNA may also play a role in the complex epigenetic interactions that occur between homologous sequences (Moazed et al, 2009). In the future, by understanding epigenetic control, researchers can uncover the research necessary to improve plant growth, yields, and transformation efficiency especially in the face of climate change and other environmental factors.</p>
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<p><img src="https://www.diagenode.com/img/areas/chromatin-and-transcription-factors.jpg" /></p>
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<h3 style="font-weight: 100; margin-top: 0;">Chromatin</h3>
<p>Chromatin consists of nucleosomes formed by a complex of histone proteins and DNA, which allows the packaging of DNA into the nucleus. The less condensed euchromatin represents transcriptionally active regions, while heterochromatin is usually inactive (Vaillant and Paszkowski, 2007). Chromatin state is known to be influenced by both DNA methylation and histone modifications which in turn impact gene expression and the structure of chromosomes. In a recent study, the role of chromatin modifications during plant reproduction elucidated 3-dimensional chromosome reorganization mediated by histones and DNA methylation (Dukowic-Schulze et al. 2017). In addition, gibberellins have been shown in increasing the level of histone acetylation, which affects regions of chromatin involved in maize seed germination (Zheng et al. 2017). Another study reports a novel function of a tomato histone deacetylase gene in the regulation of fruit ripening (Guo et al. 2017).</p>
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<p><img src="https://www.diagenode.com/img/areas/cherry-tomato-common-grape-vine-ripening-fruit-vegetable-cherry-tomatoes.jpg" /></p>
</div>
<div class="small-12 medium-9 large-9 columns">
<p>In addition, multigene families encode transcription factors, with members found throughout the genome or clustered on the same chromosome. Numerous DNA binding proteins that interact with plant promoters have been identified -- some are similar to well-characterized transcription factors in animals or yeast, while others are unique to plants. For example, diverse members of the subfamily X of the plant-specific ethylene response factor (ERF) transcription factors coordinate stress signaling with wound repair activation. Tissue repair is also enhanced through a protein complex of ERF and GRAS TFs (Heyman et. al,.2018). A compilation of known plant transcription factors can be found in the plant transcription factor database at http://plntfdb.bio.uni-potsdam.de/v3.0/.</p>
</div>
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<div class="row extra-spaced">
<div class="small-12 medium-3 large-3 columns">
<p><img src="https://www.diagenode.com/img/areas/rna-strand.jpg" /></p>
</div>
<div class="small-12 medium-9 large-9 columns">
<h3 style="font-weight: 100; margin-top: 0;">RNA</h3>
<p>Recent research shows that a number of classes of small RNAs are key epigenetic regulators. In many cases, small RNAs have been implicated in DNA methylation and chromatin modification (Meyer, 2015). In addition, the role of small RNAs has been implicated in plant stress tolerance (Kumar et al., 2017). López-Galiano et al also provided insight into a coordinated function of a miRNA gene and histone modifications in regulating the expression of a WRKY transcription factor in response to stress.</p>
<p>RNA interference (RNAi) is another epigenetic mechanism that leads to small RNA generation, which mediates gene silencing at the post-transcriptional level. RNAi technology has immense potential for plant disease resistance.</p>
</div>
</div>
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<div class="small-12 medium-3 large-3 columns">
<p><img src="https://www.diagenode.com/img/areas/dna-methylation.jpg" /></p>
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<div class="small-12 medium-9 large-9 columns">
<h3 style="font-weight: 100; margin-top: 0;">DNA methylation</h3>
<p>Plants, unlike animals, have three sites that can be methylated G, CHG (H can be A, C, T), and CHH (Law and Jacobsen, 2010). DNA methylation has attracted particular interest. In Arabidopsis, one-third of methylated genes occur in transcribed regions, and 5% of genes are methylated in promoter regions, suggesting that many of these are epigenetically regulated. (Zhang et al., 2006).</p>
<p>There are thousands of differentially methylated regions (DMRs) that influence phenotype by influencing gene expression. The analysis of epigenetic recombinant inbred line (epiRIL) plants from Arabidopsis points to the evidence of the influence of DMRs. An epiRIL results from crossing two genetically identical plants with differing DNA methylation levels (with one parent as a homozygous mutant for an essential DNA methylation maintenance gene). The offspring of these plants have similar genomes that vary only in methylation levels. Many traits have been studied using epiRILs -- flowering time, plant height, and response to abiotic stress, some of which have now been mapped to DMRs (Zhang et al. 2018)</p>
<p>Regulation by DNA methylation has been shown to be important in many aspects of plant development and response such as vernalization, hybrid vigor, and self-incompatibility (Itabashi et al. 2017). For example, vernalization treatments have shown reduced DNA methylation and subsequent initiation of flowering (Burn et al., 1993). Stress can also influence DNA methylation in plants as a response to environmental stimuli. (Steward et al., 2002; Song et al., 2012). A high degree of DNA methylation has also suggested the role in the improvement of plant fitness under different environmental conditions (Saéz-Laguna et al., 2014). In addition, methylation can affect normal fruit and hypomethylation predicts homeotic transformation and loss of fruit yield (Ong-Abdullah et al., 2015)</p>
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<p>DNA demethylation has also been implied in various aspects of plant development including pollen tube formation, embryogenesis, fruit ripening, stomatal development, and nodule formation ( Li et al. 2017). Demethylation of rice genomic DNA caused an altered pattern of gene expression, inducing dwarf plants (Sano et al., 1990).</p>
<p>Epigenetic modifications contribute to the stability and survival of the plants and their ability to adapt in different environmental conditions.</p>
</div>
</div>
<h3>Diagenode products for your epigenomics research in plants</h3>
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<h3 class="text-center"><a href="https://www.diagenode.com/en/categories/chromatin-function">Chromatin analysis</a></h3>
<center><a href="https://www.diagenode.com/en/categories/chromatin-function"><img src="https://www.diagenode.com/img/cancer/chromatin-icon.png" /></a></center>
<p class="text-left">Understand the role of chromatin in plant function and development</p>
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<li><a href="https://www.diagenode.com/en/categories/chromatin-function">Learn about our chromatin analysis products</a></li>
<li><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns"> Learn about the Universal Plant ChIP Kit</a></li>
</ul>
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<div class="small-12 medium-4 large-4 columns text-left">
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<h3 class="text-center"><a href="https://www.diagenode.com/en/categories/dna-methylation" style="color: #30415c;">DNA methylation</a></h3>
<center><a href="https://www.diagenode.com/en/categories/dna-methylation"><img src="https://www.diagenode.com/img/cancer/dna-icon.png" /></a></center>
<p class="text-left">DNA methylation and demethylation and the effects on plant response and function</p>
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<h3 class="text-center"><span class="darkgrey">Non-coding RNAs</span></h3>
<center><img src="https://www.diagenode.com/img/cancer/non-coding-icon.png" /></center>
<p class="text-left">Discover noncoding RNAs in the regulation of gene expression in plants</p>
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<li><a href="https://www.diagenode.com/en/categories/Library-preparation-for-RNA-seq">Library prep for RNA-seq studies for ncRNAs</a></li>
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<h3>References</h3>
<p><small> Burn, J. et al (1993). DNA methylation, vernalization, and the initiation of flowering. Proc. Natl. Acad. Sci. U.S.A. 90, 287–291. doi: 10.1006/scdb.1996.0055 </small></p>
<p><small> Dukowic-Schulze S, Liu C, Chen C (2017) Not just gene expression: 3D implications of chromatin modifications during sexual plant reproduction. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2222-0</small></p>
<p><small> Guo J et al (2017) A histone deacetylase gene, SlHDA3, acts as a negative regulator of fruit ripening and carotenoid accumulation. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2211-3</small></p>
<p><small> Heyman J, et.al (2018) Journal of Cell Science Emerging role of the plant ERF transcription factors in coordinating wound defense responses and repair doi: 10.1242/jcs.208215</small></p>
<p><small> Itabashi E, Osabe K, Fujimoto R, Kakizaki T (2017) Epigenetic regulation of agronomical traits in Brassicaceae. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2223-z</small></p>
<p><small> Kumar V et al (2017) Plant small RNAs: the essential epigenetic regulators of gene expression for salt-stress responses and tolerance. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2210-4</small></p>
<p><small> Law, J. A., and Jacobsen, S. E. (2010). Establishing, maintaining and modifying DNA methylation patterns in plants and animals. Nat. Rev. Genet. 11, 204–220. doi: 10.1038/nrg2719</small></p>
<p><small> Meyer, P. (2015). Epigenetic variation and environmental change. J. Exp. Bot. 66, 3541–3548. doi: 10.1093/jxb/eru502</small></p>
<p><small> Moazed, D. (2009) Small RNAs in transcriptional gene silencing and genome defence. Nature. doi: 10.1038/nature07756</small></p>
<p><small> Ong-Abdullah et al. (2015). Loss of Karma transposon methylation underlies the mantled somaclonal variant of oil palm. Nature 525, 533–537. doi: 10.1038/nature15365</small></p>
<p><small> Saéz-Laguna et al. (2014). Epigenetic variability in the genetically uniform forest tree species. PLoS One 9:e103145. doi: 10.1371/journal.pone.0103145</small></p>
<p><small> Sano, H. et al. (1990). A single treatment of rice seedlings with 5-azacytidine induces heritable dwarfism and undermethylation of genomic DNA. Mol. Gen. Genet. 220, 441–447. doi: 10.1007/BF00391751</small></p>
<p><small> Song, J et al (2012). Vernalization – A cold-induced epigenetic switch. J. Cell Sci. 125, 3723–3731. doi: 10.1242/jcs.084764</small></p>
<p><small> Steward, N et al. (2002). Periodic DNA methylation in maize nucleosomes and demethylation by environmental stress. J. Biol. Chem. 277, 37741–37746. doi: 10.1074/jbc.M204050200</small></p>
<p><small> Vaillant, I., and Paszkowski, J. (2007). Role of histone and DNA methylation in gene regulation. Curr. Opin. Plant Biol. 10, 528–533. doi: 10.1016/j.pbi.2007.06.008</small></p>
<p><small> Zhang, et al. (2006). Genome-wide high-resolution mapping and functional analysis of DNA methylation in Arabidopsis. Cell 126, 1189–1201. doi: 10.1016/j.cell.2006.08.003</small></p>
<p><small> Zhang et al. 2018 Understanding the evolutionary potential of epigenetic variation: a comparison of heritable phenotypic variation in epiRILs, RILs, and natural ecotypes of Arabidopsis thaliana. Heredity 121, 257–265 (2018) doi:10.1038/s41437-018-0095-9</small></p>
<p><small> Zheng X et al (2017) Histone acetylation is involved in GA-mediated 45S rDNA decondensation in maize aleurone layers. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2207-z</small></p>
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'
$related = array(
'id' => '2686',
'antibody_id' => null,
'name' => 'IPure kit v2',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/ipure_kit_v2_manual.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>Diagenode’s<span> </span><b>IPure</b><b><span> </span>kit<span> </span></b>is the only DNA purification kit using magnetic beads, that is specifically optimized for extracting DNA from<span> </span><b>ChIP</b><b>,<span> </span></b><b>MeDIP</b><span> </span>and<span> </span><b>CUT&Tag</b>. The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification. This simple and straightforward protocol delivers pure DNA ready for any downstream application (e.g. next generation sequencing). Comparing to phenol-chloroform extraction, the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples.</p>
<center>
<h4>High DNA recovery after purification of ChIP samples using IPure technology</h4>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-chromatin-function.png" width="500" /></center>
<p></p>
<p><small>ChIP assays were performed using different amounts of U2OS cells and the H3K9me3 antibody (Cat. No.<span> </span><span>C15410056</span>; 2 g/IP). <span>The purified DNA was eluted in 50 µl of water and quantified with a Nanodrop.</span></small></p>
<p></p>
<p><strong>Benefits of the IPure kit:</strong></p>
<ul>
<li style="text-align: left;">Provides pure DNA for any downstream application (e. g. Next generation sequencing)</li>
<li style="text-align: left;">Non-toxic</li>
<li style="text-align: left;">Fast & easy to use</li>
<li style="text-align: left;">Optimized for DNA purification after ChIP, MeDIP and CUT&Tag</li>
<li style="text-align: left;">Compatible with automation</li>
<li style="text-align: left;">Validated on the IP-Star Compact</li>
</ul>
</center>',
'label1' => 'Examples of results',
'info1' => '<h2>IPure after ChIP</h2>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-A.png" alt="ChIP-seq figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-B.png" alt="ChIP-seq figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-C.png" alt="ChIP-seq figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small><strong>Figure 1.</strong> Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors (containing the IPure module for DNA purification) and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.</small></p>
<p></p>
<h2>IPure after CUT&Tag</h2>
<p>Successful CUT&Tag results showing a low background with high region-specific enrichment has been generated using 50.000 of K562 cells, 1 µg of H3K4me3 or H3K27me3 antibody (Diagenode, C15410003 or C15410069, respectively) and proteinA-Tn5 (1:250) (Diagenode, C01070001). 1 µg of IgG (C15410206) was used as negative control. Samples were purified using the IPure kit v2 or phenol-chloroform purification. The below figures present the comparison of two purification methods.</p>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-fig2.png" style="display: block; margin-left: auto; margin-right: auto;" width="400" /></center><center>
<p style="text-align: center;"><small><strong>Figure 2.</strong> Heatmap 3kb upstream and downstream of the TSS for H3K4me3</small></p>
</center>
<p></p>
<p><img src="https://www.diagenode.com/img/product/kits/ipure-fig3.png" style="display: block; margin-left: auto; margin-right: auto;" width="600" /></p>
<p></p>
<center><small><strong>Figure 3.</strong> Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using Diagenode’s pA-Tn5 transposase (Cat. No. C01070002), H3K27me3 antibody (Cat. No. C15410069) and IPure kit v2 vs phenol chloroform purification (PC).</small></center>
<p></p>
<p></p>
<h2>IPure after MeDIP</h2>
<center><img src="https://www.diagenode.com/img/product/kits/magmedip-seq-figure_multi3.jpg" alt="medip sequencing coverage" width="600" /></center><center></center><center>
<p></p>
<small><strong>Figure 4.</strong> Consistent coverage and methylation detection from different starting amounts of DNA with the Diagenode MagMeDIP-seq Package (including the Ipure kit for DNA purification). Samples containing decreasing starting amounts of DNA (from the top down: 1000 ng (red), 250 ng (blue), 100 ng (green)) originating from human blood were prepared, revealing a consistent coverage profile for the three different starting amounts, which enables reproducible methylation detection. The CpG islands (CGIs) (marked by yellow boxes in the bottom track) are predominantly unmethylated in the human genome, and as expected, we see a depletion of reads at and around CGIs.</small></center>
<script src="chrome-extension://hhojmcideegachlhfgfdhailpfhgknjm/web_accessible_resources/index.js"></script>',
'label2' => 'iPure Workflow',
'info2' => '<h2 style="text-align: center;">Kit Method Overview & Time table</h2>
<p><img src="https://www.diagenode.com/img/product/kits/workflow-ipure-cuttag.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h3><strong>Workflow description</strong></h3>
<h5><strong>IPure after ChIP</strong></h5>
<p><strong>Step 1:</strong> Chromatin is decrosslinked and eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added.<br /> <strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet.<br /> <strong>Step 3:</strong> Proteins and remaining buffer are washed away.<br /> <strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after MeDIP</strong></h5>
<p><strong>Step 1:</strong> DNA is eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Remaining buffer are washed away.<br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after CUT&Tag</strong></h5>
<p><strong>Step 1:</strong> pA-Tn5 is inactivated and DNA released from the cells. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Proteins and remaining buffer are washed away. <br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).</p>
<p></p>
<p></p>
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<div class="small-6 columns"><center>Poplar</center><center><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></center>
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<div class="small-6 columns"><center>Tomato</center><center><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></center>
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<div class="small-6 columns"><center>Maize</center><center><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></center>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
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<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
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<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
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<div class="small-6 columns"><center>Poplar</center><center><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></center>
<p><small><strong>Figure 3.</strong> ChIP-seq was performed on Populus trichocarpa stem differenciating xylem using the Premium H3K4me3 ChIP-seq grade antibody. Libraries were prepared with the <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the input and is considered as the background enrichment. The profile in red represents enrichments along a wide region of scaffold 18. Using the same scale, the peaks of the immunoprecipitated samples are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center>Tomato</center><center><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></center>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our Premium H3K4me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
</div>
<div class="small-6 columns"><center>Maize</center><center><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></center>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our Premium H3K27me3 ChIP-seq grade antibody. Librairies were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Librairy Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<table style="width: 856px;">
<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
</td>
<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
</td>
<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
</td>
</tr>
<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9-14ac-polyclonal-antibody-classic-sample-size-10-mg">H3K9/14ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27ac-polyclonal-antibody-premium-sample-size-10-ug">H3K27ac polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
</tbody>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the trimethylated lysine 4</strong> (<strong>H3K4me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation data',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4me3</strong><br />ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K4me3 (cat. No. C15410003) and optimized PCR primer pairs for qPCR. ChIP was performed with the iDeal ChIP-seq kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p></p>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2a-ChIP-seq.jpg" width="800" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2b-ChIP-seq.jpg" width="800" /></center><center>C.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2c-ChIP-seq.jpg" width="800" /></center><center>D.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig2d-ChIP-seq.jpg" width="800" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K4me3</strong><br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 µg of the Diagenode antibody against H3K4me3 (cat. No. C15410003) as described above. The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 600 kb region of the X-chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes, respectively (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-cuttag-a.png" width="800" /></center></div>
<div class="small-12 columns"><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410003-cuttag-b.png" width="800" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K4me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 0.5 µg of the Diagenode antibody against H3K4me3 (cat. No. C15410003) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the ACTB gene on chromosome 7 (figure 3A and B, respectively).</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig3-ELISA.jpg" width="350" /></center><center></center><center></center><center></center><center></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody against H3K4me3 (cat. No. C15410003). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:11,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig4-DB.jpg" /></div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K4me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K4me3 (cat. No. C15410003), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 5A shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig5-WB.jpg" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K4me3</strong><br />Western blot was performed on whole cell extracts (40 µg, lane 1) from HeLa cells, and on 1 µg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K4me3 (cat. No. C15410003). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig6-if.jpg" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K4me3</strong><br />HeLa cells were stained with the Diagenode antibody against H3K4me3 (cat. No. C15410003) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. The cells were immunofluorescently labelled with the H3K4me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (middle), which specifically labels DNA. The right picture shows a merge of both stainings.</small></p>
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'name' => 'H3K9me3 Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone<strong> H3 containing the trimethylated lysine 9</strong> (<strong>H3K9me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9me3</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K9me3 (cat. No. C15410193) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, 2, and 5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for the heterochromatin marker Sat2 and for the ZNF510 gene, used as positive controls, and for the promoters of the active EIF4A2 and GAPDH genes, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K9me3</strong><br />ChIP was performed with 0.5 µg of the Diagenode antibody against H3K9me3 (cat. No. C15410193) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2A shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. The arrows indicate two satellite repeat regions which exhibit a stronger signal. Figures 2B, 2C and 2D show the enrichment along the ZNF510 positive control target and at the H19 and KCNQ1 imprinted genes.</small></p>
</div>
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<div class="row">
<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-CT-Fig3a.png" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15410193-CT-Fig3b.png" width="700" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K9me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K9me3 (cat. No. C15410193) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in a genomic regions on chromosome 1 containing several ZNF repeat genes and in a genomic region surrounding the KCNQ1 imprinting control gene on chromosome 11 (figure 3A and B, respectively).</small></p>
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<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-Elisa-Fig4.png" /></center></div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human H3K9me3 (cat. No. C15410193) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:87,000.</small></p>
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<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-DB-Fig5.png" /></center></div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K9me3</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9me3 (cat. No. C15410193) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest.</small></p>
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<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-WB-Fig6.png" /></center></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K9me3</strong><br />Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K9me3 (cat. No. C15410193). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15410193-IF-Fig7.png" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K9me3</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9me3 (cat. No. C15410193) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me3 antibody (middle) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of both stainings is shown on the right.</small></p>
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'description' => '<p><span>This antibody has been raised in rabbit against two KLH-conjugated synthetic peptides containing an unmodified sequence from the central part and from the C-terminus of <strong>histone H3</strong>, respectively.</span></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-chip.jpg" alt="H3pan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3pan</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3pan (Cat. No. C15310135) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 μl per ChIP experiment was analysed. IgG (2 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive MYOD1 and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-elisa.jpg" alt="H3pan Antibody ELISA validation" caption="false" width="288" height="217" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3pan (Cat. No. C15310135). The plates were coated with the peptides used for immunization. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be >1:1,000,000.</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310135-wb.jpg" alt="H3pan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against H3pan</strong><br />Whole cell extracts from HeLa cells (25 μg) were analysed by Western blot using the Diagenode antibody against H3pan (Cat. No. C15310135) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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'name' => 'H3K27me3 Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the trimethylated lysine 27</strong> (<strong>H3K27me3</strong>), using a KLH-conjugated synthetic peptide.</p>',
'label1' => 'Validation Data',
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<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig1.png" alt="H3K27me3 Antibody ChIP Grade" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2.png" alt="H3K27me3 Antibody for ChIP" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K27me3 (Cat. No. C15410195) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation. A titration consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control.</small></p>
<p><small><strong>Figure 1A.</strong> Quantitative PCR was performed with primers specific for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive TSH2B and MYT1 genes, used as positive controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
<p><small><strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me3, H3K9me3 and H3K36me3 modifications and the unmodified H3K27 as determined by qPCR. The figure clearly shows the antibody is very specific in ChIP for the H3K27me3 modification.</small></p>
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</div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2a.png" alt="H3K27me3 Antibody ChIP-seq Grade" /></p>
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<div class="extra-spaced"></div>
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<div class="extra-spaced"></div>
<div class="row">
<div class="small-12 columns">
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2b.png" alt="H3K27me3 Antibody for ChIP-seq" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2c.png" alt="H3K27me3 Antibody for ChIP-seq assay" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-ChIP-Fig2d.png" alt="H3K27me3 Antibody validated in ChIP-seq" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 µg of the Diagenode antibody against H3K27me3 (Cat. No. C15410195) as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment in genomic regions of chromosome 6 and 20, surrounding the TSH2B and MYT1 positive control genes (fig 2A and 2B, respectively), and in two genomic regions of chromosome 1 and X (figure 2C and D).</small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
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<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-CUTTAG-Fig3A.png" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410195-CUTTAG-Fig3B.png" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K27me3</strong><br />CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K27me3 (cat. No. C15410195) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions on chromosome and 13 and 20 (figure 3A and B, respectively).</small></p>
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<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-ELISA-Fig4.png" alt="H3K27me3 Antibody ELISA Validation " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me3 (Cat. No. C15410195). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:3,000.</small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-DB-Fig5a.png" alt="H3K27me3 Antibody Dot Blot Validation " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K27me3</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3 (Cat. No. C15410195) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 5 shows a high specificity of the antibody for the modification of interest. Please note that the antibody also recognizes the modification if S28 is phosphorylated.</small></p>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-WB-Fig6.png" alt="H3K27me3 Antibody validated in Western Blot" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 6. Western blot analysis using the Diagenode antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell (25 µg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells, and on 1 µg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H3K27me3 (cat. No. C15410195) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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</div>
<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410195-IF-Fig7.png" alt="H3K27me3 Antibody validated for Immunofluorescence" /></p>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 7. Immunofluorescence using the Diagenode antibody directed against H3K27me3</strong><br />Human HeLa cells were stained with the Diagenode antibody against H3K27me3 (Cat. No. C15410195) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p style="text-align: justify;"><span>Previous name of the kit: Chromatin Shearing Optimization Kit (Universal Plant ChIP-seq kit)<br /></span></p>
<p style="text-align: justify;"><span>The first critical step of a successful ChIP experiment is the best preparation of sheared chromatin. This <strong>Chromatin EasyShear Kit</strong> is designed to be used in conjunction with the <strong>Universal Plant ChIP-seq kit</strong> and contains the right level of <strong>detergent</strong> for extraction of highest quality plant chromatin for ChIP. In addition, the signature</span><span> crosslinking containers of this kit provide a simple and reliable method for fixation. The content of this kit is enough to perform 12 chromatin extractions.<br /></span></p>
<p style="text-align: justify;"><span>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
<p style="text-align: justify;"><span>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></span></p>',
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'meta_description' => 'Chromatin Shearing Optimization Kit designed to be used in conjunction with the Universal Plant ChIP-seq kit, providing high quality plant chromatin for ChIP',
'modified' => '2023-04-20 16:17:53',
'created' => '2016-02-22 15:08:23',
'ProductsRelated' => array(
[maximum depth reached]
),
'Image' => array([maximum depth reached])
),
(int) 5 => array(
'id' => '2791',
'antibody_id' => null,
'name' => 'Universal Plant ChIP-seq kit',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/Universal_Plant_ChIPseq-kit-manual.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p style="text-align: justify;">The <strong>Universal Plant ChIP-seq kit</strong> offers the convenience of extracting plant chromatin from a wide variety of plants including Arabidopsis, maize, rice, tomato and poplar. This complete kit has been specifically optimized for <strong>plant chromatin extraction</strong> and includes reagents for chromatin preparation, immunoprecipitation, plant-specific control primer pairs, control antibody, and DNA purification.</p>',
'label1' => 'Characteristics',
'info1' => '<ul>
<li><strong>Universal compatiblity</strong> with a wide variety of plant species</li>
<li>Optimized and <strong>complete kit</strong> for start-to-finish plant ChIP</li>
<li>Includes <strong>plant-specific control</strong> primers and control antibody<strong></strong></li>
</ul>
<h3>Successful ChIP-seq experiments for a variety of plants</h3>
<div class="row">
<div class="small-6 columns">
<h4 class="text-center">Arabidopsis</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG1"> <img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A-small.jpg" /> </a></p>
<div id="IMG1" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/Plant-ChIP-figure-3-A.png" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 1.</strong> ChIP-seq was performed on Arabidopsis thaliana (Col-0) seedlings using our <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng (green), 500 pg (orange) and 100 pg (red) IP'd DNA and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a public dataset (NCBI GEO Dataset GSM1193621) that we used as an external reference. Enrichments along a wide region of chromosome 5 are uniform regardless of the starting material amount for the preparation of the library.</small></p>
</div>
<div class="small-6 columns">
<h4 class="text-center">Poplar</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG2"><img src="https://www.diagenode.com/img/landing-pages/poplar-small.jpg" /> </a></p>
<div id="IMG2" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/poplar.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 3.</strong> ChIP-seq was performed on Populus trichocarpa stem differenciating xylem using the <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with the <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the input and is considered as the background enrichment. The profile in red represents enrichments along a wide region of scaffold 18. Using the same scale, the peaks of the immunoprecipitated samples are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<h4 class="text-center">Tomato</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG3"> <img src="https://www.diagenode.com/img/landing-pages/tomtato-small.jpg" /> </a></p>
<div id="IMG3" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/tomtato.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 2.</strong> ChIP-seq was performed on Solanum lycopersicum cv. Micro-Tom young leaves using our <a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">Premium H3K4me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 750 pg of immunoprecipitated DNA using the Universal Plant ChIP-seq kit (red) and sequenced on an Illumina® HiSeq 2500. The enrichment in blue represents a dataset obtained from Nguyen et al. 2014 that we used as an external reference. Enrichments are higher and consistent with the reference data along a wide region of chromosome 1.</small></p>
</div>
<div class="small-6 columns">
<h4 class="text-center">Maize</h4>
<p class="text-center"><a href="#" data-reveal-id="IMG4"> <img src="https://www.diagenode.com/img/landing-pages/maize-small.jpg" /> </a></p>
<div id="IMG4" class="reveal-modal" data-reveal="" aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
<p class="text-center"><img src="https://www.diagenode.com/img/landing-pages/maize.jpg" /></p>
<a class="close-reveal-modal" aria-label="Close">×</a></div>
<p><small><strong>Figure 4.</strong> ChIP-seq was performed on Zea mays cv. B73 inner stem using our <a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">Premium H3K27me3 ChIP-seq grade antibody</a>. Libraries were prepared with our <a href="https://www.diagenode.com/p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> from 1 ng of immunoprecipitated DNA using the Universal Plant ChIP-seq kit and 1 ng of Input and sequenced on an Illumina® HiSeq 2500. The enrichment in green represents the Input and is considered as the background enrichment. The enrichment in red represents enrichments along a wide region of chromosome 3. Using the same scale, the peaks of the immunoprecipitated sample are significantly higher than those of the input, indicating a successful ChIP-seq experiment.</small></p>
</div>
</div>
<p><strong> </strong></p>
<table style="width: 856px;">
<tbody>
<tr>
<td style="width: 224px;">
<h4><strong>Plant Species</strong></h4>
</td>
<td style="width: 341px;">
<h4><strong>Validated antibodies</strong></h4>
</td>
<td style="width: 357px;">
<h4><strong>Validated primer pairs</strong></h4>
</td>
</tr>
<tr>
<td style="width: 224px;"><strong>Arabidopsis (<em>Arabidopsis thaliana</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-actin-atg-primer-pair-50-ul">Arabidopsis Actin ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-monoclonal-antibody-classic-50-ug-50-ul">H3K4me3 monoclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-atg-primer-pair-50-ul">Arabidopsis FLC-ATG primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/arabidopsis-flc-intron1-primer-pair-50-ul">Arabidopsis FLC-intron1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me3-polyclonal-antibody-classic-sample-size-10-ug">H3K9me3 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9-14ac-polyclonal-antibody-classic-sample-size-10-mg">H3K9/14ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27ac-polyclonal-antibody-premium-sample-size-10-ug">H3K27ac polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Maize (<em>Zea mays</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/maize-B73-inner-stem-ZmB1-UTR-primer-pair-50ul">Maize B73 inner stem ZmB1-UTR primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/Maize-B73-inner-stem-ZmCopia-primer-pair-50ul">Maize B73 inner stem ZmCopia primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Tomato (<em>Solanum lycopersicum</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr2-reg8-primer-pair-50ul">Tomato leaves SlChr2-reg8 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/tomato-leaves-SlChr4-NC1-primer-pair-50ul">Tomato leaves SlChr4-NC1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Rice (<em>Oriza sativa</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsChr4-reg9-primer-pair-50ul">Rice seedlings OsChr4-reg9 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9me2-polyclonal-antibody-classic-50-ug-44-ul">H3K9me2 polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/rice-seedlings-OsMADS6-primer-pair-50ul">Rice seedlings OsMADS6 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k36me3-polyclonal-antibody-premium-sample-size-10-ug">H3K36me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"><strong>Poplar (<em>Populus trichocarpa, Populus tremula x alba</em>)</strong></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k4me3-polyclonal-antibody-premium-sample-size-10-ug">H3K4me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrCopia-orth-primer-pair-50ul">Poplar xylem PtrCopia-orth primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k9ac-polyclonal-antibody-classic-sample-size-10-ug">H3K9ac polyclonal antibody - Classic</a></td>
<td style="width: 357px;"><a href="https://www.diagenode.com/p/poplar-xylem-PtrMYBTF1-primer-pair-50ul">Poplar xylem PtrMYBTF1 primer pair</a></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3pan-monoclonal-antibody-classic-50-mg-100-ml">H3pan monoclonal antibody - Classic</a></td>
<td style="width: 357px;"></td>
</tr>
<tr>
<td style="width: 224px;"></td>
<td style="width: 341px;"><a href="https://www.diagenode.com/p/h3k27me3-polyclonal-antibody-premium-sample-size-10-ug">H3K27me3 polyclonal antibody - Premium</a></td>
<td style="width: 357px;"></td>
</tr>
</tbody>
</table>
<p><strong></strong></p>
<p><strong></strong></p>',
'label2' => '',
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'format' => '24 rxns',
'catalog_number' => 'C01010152',
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'sf_code' => 'C01010152-',
'type' => 'RFR',
'search_order' => '04-undefined',
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'slug' => 'universal-plant-chip-seq-kit-x24-24-rxns',
'meta_title' => 'Universal Plant ChIP-seq kit | Diagenode',
'meta_keywords' => 'plant epigenetics, plant ChIP, plant ChIP-seq, Arabidopsis, maize, rice, tomato, poplar',
'meta_description' => 'Optimized extraction of plant chromatin from Arabidopsis,maize,rice,tomato,poplar.Complete ChIP kit including plant-specific control primer pairs and antibody',
'modified' => '2023-04-20 16:15:50',
'created' => '2016-02-22 15:08:28',
'ProductsRelated' => array(
[maximum depth reached]
),
'Image' => array(
[maximum depth reached]
)
),
(int) 6 => array(
'id' => '2686',
'antibody_id' => null,
'name' => 'IPure kit v2',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/ipure_kit_v2_manual.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>Diagenode’s<span> </span><b>IPure</b><b><span> </span>kit<span> </span></b>is the only DNA purification kit using magnetic beads, that is specifically optimized for extracting DNA from<span> </span><b>ChIP</b><b>,<span> </span></b><b>MeDIP</b><span> </span>and<span> </span><b>CUT&Tag</b>. The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification. This simple and straightforward protocol delivers pure DNA ready for any downstream application (e.g. next generation sequencing). Comparing to phenol-chloroform extraction, the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples.</p>
<center>
<h4>High DNA recovery after purification of ChIP samples using IPure technology</h4>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-chromatin-function.png" width="500" /></center>
<p></p>
<p><small>ChIP assays were performed using different amounts of U2OS cells and the H3K9me3 antibody (Cat. No.<span> </span><span>C15410056</span>; 2 g/IP). <span>The purified DNA was eluted in 50 µl of water and quantified with a Nanodrop.</span></small></p>
<p></p>
<p><strong>Benefits of the IPure kit:</strong></p>
<ul>
<li style="text-align: left;">Provides pure DNA for any downstream application (e. g. Next generation sequencing)</li>
<li style="text-align: left;">Non-toxic</li>
<li style="text-align: left;">Fast & easy to use</li>
<li style="text-align: left;">Optimized for DNA purification after ChIP, MeDIP and CUT&Tag</li>
<li style="text-align: left;">Compatible with automation</li>
<li style="text-align: left;">Validated on the IP-Star Compact</li>
</ul>
</center>',
'label1' => 'Examples of results',
'info1' => '<h2>IPure after ChIP</h2>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-A.png" alt="ChIP-seq figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-B.png" alt="ChIP-seq figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-C.png" alt="ChIP-seq figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small><strong>Figure 1.</strong> Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors (containing the IPure module for DNA purification) and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.</small></p>
<p></p>
<h2>IPure after CUT&Tag</h2>
<p>Successful CUT&Tag results showing a low background with high region-specific enrichment has been generated using 50.000 of K562 cells, 1 µg of H3K4me3 or H3K27me3 antibody (Diagenode, C15410003 or C15410069, respectively) and proteinA-Tn5 (1:250) (Diagenode, C01070001). 1 µg of IgG (C15410206) was used as negative control. Samples were purified using the IPure kit v2 or phenol-chloroform purification. The below figures present the comparison of two purification methods.</p>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-fig2.png" style="display: block; margin-left: auto; margin-right: auto;" width="400" /></center><center>
<p style="text-align: center;"><small><strong>Figure 2.</strong> Heatmap 3kb upstream and downstream of the TSS for H3K4me3</small></p>
</center>
<p></p>
<p><img src="https://www.diagenode.com/img/product/kits/ipure-fig3.png" style="display: block; margin-left: auto; margin-right: auto;" width="600" /></p>
<p></p>
<center><small><strong>Figure 3.</strong> Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using Diagenode’s pA-Tn5 transposase (Cat. No. C01070002), H3K27me3 antibody (Cat. No. C15410069) and IPure kit v2 vs phenol chloroform purification (PC).</small></center>
<p></p>
<p></p>
<h2>IPure after MeDIP</h2>
<center><img src="https://www.diagenode.com/img/product/kits/magmedip-seq-figure_multi3.jpg" alt="medip sequencing coverage" width="600" /></center><center></center><center>
<p></p>
<small><strong>Figure 4.</strong> Consistent coverage and methylation detection from different starting amounts of DNA with the Diagenode MagMeDIP-seq Package (including the Ipure kit for DNA purification). Samples containing decreasing starting amounts of DNA (from the top down: 1000 ng (red), 250 ng (blue), 100 ng (green)) originating from human blood were prepared, revealing a consistent coverage profile for the three different starting amounts, which enables reproducible methylation detection. The CpG islands (CGIs) (marked by yellow boxes in the bottom track) are predominantly unmethylated in the human genome, and as expected, we see a depletion of reads at and around CGIs.</small></center>
<script src="chrome-extension://hhojmcideegachlhfgfdhailpfhgknjm/web_accessible_resources/index.js"></script>',
'label2' => 'iPure Workflow',
'info2' => '<h2 style="text-align: center;">Kit Method Overview & Time table</h2>
<p><img src="https://www.diagenode.com/img/product/kits/workflow-ipure-cuttag.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h3><strong>Workflow description</strong></h3>
<h5><strong>IPure after ChIP</strong></h5>
<p><strong>Step 1:</strong> Chromatin is decrosslinked and eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added.<br /> <strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet.<br /> <strong>Step 3:</strong> Proteins and remaining buffer are washed away.<br /> <strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after MeDIP</strong></h5>
<p><strong>Step 1:</strong> DNA is eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Remaining buffer are washed away.<br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after CUT&Tag</strong></h5>
<p><strong>Step 1:</strong> pA-Tn5 is inactivated and DNA released from the cells. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Proteins and remaining buffer are washed away. <br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).</p>
<p></p>
<p></p>
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<div class="large-12 columns">Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on a genome-wide scale. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</div>
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<h5 class="large-12 columns"><strong></strong></h5>
<h5 class="large-12 columns"><strong>The ChIP-seq workflow</strong></h5>
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<li class="large-12 columns"><strong>Chromatin preparation: </strong>Crosslink chromatin-bound proteins (histones or transcription factors) to DNA followed by cell lysis.</li>
<li class="large-12 columns"><strong>Chromatin shearing:</strong> Fragment chromatin by sonication to desired fragment size (100-500 bp)</li>
<li class="large-12 columns"><strong>Chromatin IP</strong>: Capture protein-DNA complexes with <strong><a href="../categories/chip-seq-grade-antibodies">specific ChIP-seq grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: Reverse cross-links, elute, and purify </li>
<li class="large-12 columns"><strong>NGS Library Preparation</strong>: Ligate adapters and amplify IP'd material</li>
<li class="large-12 columns"><strong>Bioinformatic analysis</strong>: Perform r<span style="font-weight: 400;">ead filtering and trimming</span>, r<span style="font-weight: 400;">ead specific alignment, enrichment specific peak calling, QC metrics, multi-sample cross-comparison etc. </span></li>
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<h3 class="text-center" style="color: #b21329;">Need guidance?</h3>
<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
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<p class="text-justify">Chromatin Immunoprecipitation (ChIP) coupled with quantitative PCR can be used to investigate protein-DNA interaction at known genomic binding sites. if sites are not known, qPCR primers can also be designed against potential regulatory regions such as promoters. ChIP-qPCR is advantageous in studies that focus on specific genes and potential regulatory regions across differing experimental conditions as the cost of performing real-time PCR is minimal. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</p>
<p class="text-justify"><strong>The ChIP-qPCR workflow</strong></p>
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<li class="large-12 columns"><strong>Chromatin IP</strong>: protein-DNA complexe capture using<strong> <a href="https://www.diagenode.com/en/categories/chip-grade-antibodies">specific ChIP-grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: chromatin reverse cross-linking and elution followed by purification<strong> </strong></li>
<li class="large-12 columns"><strong>qPCR and analysis</strong>: using previously designed primers to amplify IP'd material at specific loci</li>
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<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
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<p><img src="https://www.diagenode.com/img/areas/plant.jpg" /></p>
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<div class="extra-spaced">
<h2>Epigenetic Regulation in Plants</h2>
<p>Plants utilize a number of gene regulation mechanisms to ensure proper development, function, growth, and survival under different environmental conditions. Plants depend on changes in gene expression to respond to environmental stimuli, in which the full repertoire of histone modifications, DNA methylation, and small ncRNAs play an important role in epigenetic regulation.</p>
<p>Studying the epigenetics of model plants such as Arabidopsis thaliana have allowed researchers to understand pathways that maintain chromatin modifications as well as the mapping of modifications such as DNA methylation on a genome-wide scale. Small RNAs have also been implicated in playing a role in the distribution of chromatin modifications, and RNA may also play a role in the complex epigenetic interactions that occur between homologous sequences (Moazed et al, 2009). In the future, by understanding epigenetic control, researchers can uncover the research necessary to improve plant growth, yields, and transformation efficiency especially in the face of climate change and other environmental factors.</p>
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<div class="row extra-spaced">
<div class="small-12 medium-3 large-3 columns">
<p><img src="https://www.diagenode.com/img/areas/chromatin-and-transcription-factors.jpg" /></p>
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<div class="small-12 medium-9 large-9 columns">
<h3 style="font-weight: 100; margin-top: 0;">Chromatin</h3>
<p>Chromatin consists of nucleosomes formed by a complex of histone proteins and DNA, which allows the packaging of DNA into the nucleus. The less condensed euchromatin represents transcriptionally active regions, while heterochromatin is usually inactive (Vaillant and Paszkowski, 2007). Chromatin state is known to be influenced by both DNA methylation and histone modifications which in turn impact gene expression and the structure of chromosomes. In a recent study, the role of chromatin modifications during plant reproduction elucidated 3-dimensional chromosome reorganization mediated by histones and DNA methylation (Dukowic-Schulze et al. 2017). In addition, gibberellins have been shown in increasing the level of histone acetylation, which affects regions of chromatin involved in maize seed germination (Zheng et al. 2017). Another study reports a novel function of a tomato histone deacetylase gene in the regulation of fruit ripening (Guo et al. 2017).</p>
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<div class="small-12 medium-3 large-3 columns">
<p><img src="https://www.diagenode.com/img/areas/cherry-tomato-common-grape-vine-ripening-fruit-vegetable-cherry-tomatoes.jpg" /></p>
</div>
<div class="small-12 medium-9 large-9 columns">
<p>In addition, multigene families encode transcription factors, with members found throughout the genome or clustered on the same chromosome. Numerous DNA binding proteins that interact with plant promoters have been identified -- some are similar to well-characterized transcription factors in animals or yeast, while others are unique to plants. For example, diverse members of the subfamily X of the plant-specific ethylene response factor (ERF) transcription factors coordinate stress signaling with wound repair activation. Tissue repair is also enhanced through a protein complex of ERF and GRAS TFs (Heyman et. al,.2018). A compilation of known plant transcription factors can be found in the plant transcription factor database at http://plntfdb.bio.uni-potsdam.de/v3.0/.</p>
</div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-3 large-3 columns">
<p><img src="https://www.diagenode.com/img/areas/rna-strand.jpg" /></p>
</div>
<div class="small-12 medium-9 large-9 columns">
<h3 style="font-weight: 100; margin-top: 0;">RNA</h3>
<p>Recent research shows that a number of classes of small RNAs are key epigenetic regulators. In many cases, small RNAs have been implicated in DNA methylation and chromatin modification (Meyer, 2015). In addition, the role of small RNAs has been implicated in plant stress tolerance (Kumar et al., 2017). López-Galiano et al also provided insight into a coordinated function of a miRNA gene and histone modifications in regulating the expression of a WRKY transcription factor in response to stress.</p>
<p>RNA interference (RNAi) is another epigenetic mechanism that leads to small RNA generation, which mediates gene silencing at the post-transcriptional level. RNAi technology has immense potential for plant disease resistance.</p>
</div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-3 large-3 columns">
<p><img src="https://www.diagenode.com/img/areas/dna-methylation.jpg" /></p>
</div>
<div class="small-12 medium-9 large-9 columns">
<h3 style="font-weight: 100; margin-top: 0;">DNA methylation</h3>
<p>Plants, unlike animals, have three sites that can be methylated G, CHG (H can be A, C, T), and CHH (Law and Jacobsen, 2010). DNA methylation has attracted particular interest. In Arabidopsis, one-third of methylated genes occur in transcribed regions, and 5% of genes are methylated in promoter regions, suggesting that many of these are epigenetically regulated. (Zhang et al., 2006).</p>
<p>There are thousands of differentially methylated regions (DMRs) that influence phenotype by influencing gene expression. The analysis of epigenetic recombinant inbred line (epiRIL) plants from Arabidopsis points to the evidence of the influence of DMRs. An epiRIL results from crossing two genetically identical plants with differing DNA methylation levels (with one parent as a homozygous mutant for an essential DNA methylation maintenance gene). The offspring of these plants have similar genomes that vary only in methylation levels. Many traits have been studied using epiRILs -- flowering time, plant height, and response to abiotic stress, some of which have now been mapped to DMRs (Zhang et al. 2018)</p>
<p>Regulation by DNA methylation has been shown to be important in many aspects of plant development and response such as vernalization, hybrid vigor, and self-incompatibility (Itabashi et al. 2017). For example, vernalization treatments have shown reduced DNA methylation and subsequent initiation of flowering (Burn et al., 1993). Stress can also influence DNA methylation in plants as a response to environmental stimuli. (Steward et al., 2002; Song et al., 2012). A high degree of DNA methylation has also suggested the role in the improvement of plant fitness under different environmental conditions (Saéz-Laguna et al., 2014). In addition, methylation can affect normal fruit and hypomethylation predicts homeotic transformation and loss of fruit yield (Ong-Abdullah et al., 2015)</p>
</div>
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<p><img src="https://www.diagenode.com/img/areas/plant-development.jpg" class="left" style="padding-right: 15px;" /></p>
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<div class="small-12 medium-9 large-9 columns">
<p>DNA demethylation has also been implied in various aspects of plant development including pollen tube formation, embryogenesis, fruit ripening, stomatal development, and nodule formation ( Li et al. 2017). Demethylation of rice genomic DNA caused an altered pattern of gene expression, inducing dwarf plants (Sano et al., 1990).</p>
<p>Epigenetic modifications contribute to the stability and survival of the plants and their ability to adapt in different environmental conditions.</p>
</div>
</div>
<h3>Diagenode products for your epigenomics research in plants</h3>
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<div class="small-12 medium-4 large-4 columns text-left">
<div class="panel" style="border-color: #099f92; height: 275px;">
<h3 class="text-center"><a href="https://www.diagenode.com/en/categories/chromatin-function">Chromatin analysis</a></h3>
<center><a href="https://www.diagenode.com/en/categories/chromatin-function"><img src="https://www.diagenode.com/img/cancer/chromatin-icon.png" /></a></center>
<p class="text-left">Understand the role of chromatin in plant function and development</p>
</div>
<ul>
<li><a href="https://www.diagenode.com/en/categories/chromatin-function">Learn about our chromatin analysis products</a></li>
<li><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns"> Learn about the Universal Plant ChIP Kit</a></li>
</ul>
</div>
<div class="small-12 medium-4 large-4 columns text-left">
<div class="panel" style="border-color: #30415c; height: 275px;">
<h3 class="text-center"><a href="https://www.diagenode.com/en/categories/dna-methylation" style="color: #30415c;">DNA methylation</a></h3>
<center><a href="https://www.diagenode.com/en/categories/dna-methylation"><img src="https://www.diagenode.com/img/cancer/dna-icon.png" /></a></center>
<p class="text-left">DNA methylation and demethylation and the effects on plant response and function</p>
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<li><a href="https://www.diagenode.com/en/categories/dna-methylation">Discover DNA methylation analysis solutions at any resolution</a></li>
</ul>
</div>
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<div class="panel" style="border-color: #474546; height: 275px;">
<h3 class="text-center"><span class="darkgrey">Non-coding RNAs</span></h3>
<center><img src="https://www.diagenode.com/img/cancer/non-coding-icon.png" /></center>
<p class="text-left">Discover noncoding RNAs in the regulation of gene expression in plants</p>
</div>
<ul>
<li><a href="https://www.diagenode.com/en/categories/Library-preparation-for-RNA-seq">Library prep for RNA-seq studies for ncRNAs</a></li>
</ul>
</div>
</div>
<h3>References</h3>
<p><small> Burn, J. et al (1993). DNA methylation, vernalization, and the initiation of flowering. Proc. Natl. Acad. Sci. U.S.A. 90, 287–291. doi: 10.1006/scdb.1996.0055 </small></p>
<p><small> Dukowic-Schulze S, Liu C, Chen C (2017) Not just gene expression: 3D implications of chromatin modifications during sexual plant reproduction. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2222-0</small></p>
<p><small> Guo J et al (2017) A histone deacetylase gene, SlHDA3, acts as a negative regulator of fruit ripening and carotenoid accumulation. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2211-3</small></p>
<p><small> Heyman J, et.al (2018) Journal of Cell Science Emerging role of the plant ERF transcription factors in coordinating wound defense responses and repair doi: 10.1242/jcs.208215</small></p>
<p><small> Itabashi E, Osabe K, Fujimoto R, Kakizaki T (2017) Epigenetic regulation of agronomical traits in Brassicaceae. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2223-z</small></p>
<p><small> Kumar V et al (2017) Plant small RNAs: the essential epigenetic regulators of gene expression for salt-stress responses and tolerance. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2210-4</small></p>
<p><small> Law, J. A., and Jacobsen, S. E. (2010). Establishing, maintaining and modifying DNA methylation patterns in plants and animals. Nat. Rev. Genet. 11, 204–220. doi: 10.1038/nrg2719</small></p>
<p><small> Meyer, P. (2015). Epigenetic variation and environmental change. J. Exp. Bot. 66, 3541–3548. doi: 10.1093/jxb/eru502</small></p>
<p><small> Moazed, D. (2009) Small RNAs in transcriptional gene silencing and genome defence. Nature. doi: 10.1038/nature07756</small></p>
<p><small> Ong-Abdullah et al. (2015). Loss of Karma transposon methylation underlies the mantled somaclonal variant of oil palm. Nature 525, 533–537. doi: 10.1038/nature15365</small></p>
<p><small> Saéz-Laguna et al. (2014). Epigenetic variability in the genetically uniform forest tree species. PLoS One 9:e103145. doi: 10.1371/journal.pone.0103145</small></p>
<p><small> Sano, H. et al. (1990). A single treatment of rice seedlings with 5-azacytidine induces heritable dwarfism and undermethylation of genomic DNA. Mol. Gen. Genet. 220, 441–447. doi: 10.1007/BF00391751</small></p>
<p><small> Song, J et al (2012). Vernalization – A cold-induced epigenetic switch. J. Cell Sci. 125, 3723–3731. doi: 10.1242/jcs.084764</small></p>
<p><small> Steward, N et al. (2002). Periodic DNA methylation in maize nucleosomes and demethylation by environmental stress. J. Biol. Chem. 277, 37741–37746. doi: 10.1074/jbc.M204050200</small></p>
<p><small> Vaillant, I., and Paszkowski, J. (2007). Role of histone and DNA methylation in gene regulation. Curr. Opin. Plant Biol. 10, 528–533. doi: 10.1016/j.pbi.2007.06.008</small></p>
<p><small> Zhang, et al. (2006). Genome-wide high-resolution mapping and functional analysis of DNA methylation in Arabidopsis. Cell 126, 1189–1201. doi: 10.1016/j.cell.2006.08.003</small></p>
<p><small> Zhang et al. 2018 Understanding the evolutionary potential of epigenetic variation: a comparison of heritable phenotypic variation in epiRILs, RILs, and natural ecotypes of Arabidopsis thaliana. Heredity 121, 257–265 (2018) doi:10.1038/s41437-018-0095-9</small></p>
<p><small> Zheng X et al (2017) Histone acetylation is involved in GA-mediated 45S rDNA decondensation in maize aleurone layers. Plant Cell Rep. https://dx.doi.org/10.1007/s00299-017-2207-z</small></p>
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(int) 7 => 'FI',
(int) 8 => 'NL',
(int) 9 => 'BE',
(int) 10 => 'LU',
(int) 11 => 'FR',
(int) 12 => 'DE',
(int) 13 => 'CH',
(int) 14 => 'AT',
(int) 15 => 'ES',
(int) 16 => 'IT',
(int) 17 => 'PT'
)
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'
$related = array(
'id' => '2686',
'antibody_id' => null,
'name' => 'IPure kit v2',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/ipure_kit_v2_manual.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>Diagenode’s<span> </span><b>IPure</b><b><span> </span>kit<span> </span></b>is the only DNA purification kit using magnetic beads, that is specifically optimized for extracting DNA from<span> </span><b>ChIP</b><b>,<span> </span></b><b>MeDIP</b><span> </span>and<span> </span><b>CUT&Tag</b>. The use of the magnetic beads allows for a clear separation of DNA and increases therefore the reproducibility of your DNA purification. This simple and straightforward protocol delivers pure DNA ready for any downstream application (e.g. next generation sequencing). Comparing to phenol-chloroform extraction, the IPure technology has the advantage of being nontoxic and much easier to be carried out on multiple samples.</p>
<center>
<h4>High DNA recovery after purification of ChIP samples using IPure technology</h4>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-chromatin-function.png" width="500" /></center>
<p></p>
<p><small>ChIP assays were performed using different amounts of U2OS cells and the H3K9me3 antibody (Cat. No.<span> </span><span>C15410056</span>; 2 g/IP). <span>The purified DNA was eluted in 50 µl of water and quantified with a Nanodrop.</span></small></p>
<p></p>
<p><strong>Benefits of the IPure kit:</strong></p>
<ul>
<li style="text-align: left;">Provides pure DNA for any downstream application (e. g. Next generation sequencing)</li>
<li style="text-align: left;">Non-toxic</li>
<li style="text-align: left;">Fast & easy to use</li>
<li style="text-align: left;">Optimized for DNA purification after ChIP, MeDIP and CUT&Tag</li>
<li style="text-align: left;">Compatible with automation</li>
<li style="text-align: left;">Validated on the IP-Star Compact</li>
</ul>
</center>',
'label1' => 'Examples of results',
'info1' => '<h2>IPure after ChIP</h2>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-A.png" alt="ChIP-seq figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-B.png" alt="ChIP-seq figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/ideal-TF-chip-seq-C.png" alt="ChIP-seq figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small><strong>Figure 1.</strong> Chromatin Immunoprecipitation has been performed using chromatin from HeLa cells, the iDeal ChIP-seq kit for Transcription Factors (containing the IPure module for DNA purification) and the Diagenode ChIP-seq-grade HDAC1 (A), LSD1 (B) and p53 antibody (C). The IP'd DNA was subsequently analysed on an Illumina® Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. This figure shows the peak distribution in regions of chromosome 3 (A), chromosome 12 (B) and chromosome 6 (C) respectively.</small></p>
<p></p>
<h2>IPure after CUT&Tag</h2>
<p>Successful CUT&Tag results showing a low background with high region-specific enrichment has been generated using 50.000 of K562 cells, 1 µg of H3K4me3 or H3K27me3 antibody (Diagenode, C15410003 or C15410069, respectively) and proteinA-Tn5 (1:250) (Diagenode, C01070001). 1 µg of IgG (C15410206) was used as negative control. Samples were purified using the IPure kit v2 or phenol-chloroform purification. The below figures present the comparison of two purification methods.</p>
<center><img src="https://www.diagenode.com/img/product/kits/ipure-fig2.png" style="display: block; margin-left: auto; margin-right: auto;" width="400" /></center><center>
<p style="text-align: center;"><small><strong>Figure 2.</strong> Heatmap 3kb upstream and downstream of the TSS for H3K4me3</small></p>
</center>
<p></p>
<p><img src="https://www.diagenode.com/img/product/kits/ipure-fig3.png" style="display: block; margin-left: auto; margin-right: auto;" width="600" /></p>
<p></p>
<center><small><strong>Figure 3.</strong> Integrative genomics viewer (IGV) visualization of CUT&Tag experiments using Diagenode’s pA-Tn5 transposase (Cat. No. C01070002), H3K27me3 antibody (Cat. No. C15410069) and IPure kit v2 vs phenol chloroform purification (PC).</small></center>
<p></p>
<p></p>
<h2>IPure after MeDIP</h2>
<center><img src="https://www.diagenode.com/img/product/kits/magmedip-seq-figure_multi3.jpg" alt="medip sequencing coverage" width="600" /></center><center></center><center>
<p></p>
<small><strong>Figure 4.</strong> Consistent coverage and methylation detection from different starting amounts of DNA with the Diagenode MagMeDIP-seq Package (including the Ipure kit for DNA purification). Samples containing decreasing starting amounts of DNA (from the top down: 1000 ng (red), 250 ng (blue), 100 ng (green)) originating from human blood were prepared, revealing a consistent coverage profile for the three different starting amounts, which enables reproducible methylation detection. The CpG islands (CGIs) (marked by yellow boxes in the bottom track) are predominantly unmethylated in the human genome, and as expected, we see a depletion of reads at and around CGIs.</small></center>
<script src="chrome-extension://hhojmcideegachlhfgfdhailpfhgknjm/web_accessible_resources/index.js"></script>',
'label2' => 'iPure Workflow',
'info2' => '<h2 style="text-align: center;">Kit Method Overview & Time table</h2>
<p><img src="https://www.diagenode.com/img/product/kits/workflow-ipure-cuttag.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h3><strong>Workflow description</strong></h3>
<h5><strong>IPure after ChIP</strong></h5>
<p><strong>Step 1:</strong> Chromatin is decrosslinked and eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added.<br /> <strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet.<br /> <strong>Step 3:</strong> Proteins and remaining buffer are washed away.<br /> <strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after MeDIP</strong></h5>
<p><strong>Step 1:</strong> DNA is eluted from beads (magnetic or agarose) which are discarded. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Remaining buffer are washed away.<br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).<br /><br /><br /></p>
<h5><strong>IPure after CUT&Tag</strong></h5>
<p><strong>Step 1:</strong> pA-Tn5 is inactivated and DNA released from the cells. <strong>Magnetic beads</strong> <strong>for purification</strong> are added. <br /><strong>Step 2:</strong> Magnetic beads acquire positive charge to bind the negatively charged phosphate backbone of DNA. DNA-bead complex is separated using a magnet. <br /><strong>Step 3:</strong> Proteins and remaining buffer are washed away. <br /><strong>Step 4:</strong> DNA is eluted from magnetic beads, which are discarded. Purified DNA is ready for any downstream application (NGS, qPCR, amplification, microarray).</p>
<p></p>
<p></p>
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