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'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the trimethylated lysine 36 (H3K36me3), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2.png" alt="H3K36me3 Antibody ChIP Grade" caption="false" width="432" height="453" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 µl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 µg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with 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 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes </small></p>
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<div class="row">
<div class="small-12 columns">
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B2.png" alt="H3K36me3 Antibody ChIP-seq Grade" caption="false" width="700" height="128" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B3.png" alt="H3K36me3 Antibody for ChIP-seq" caption="false" width="700" height="107" /></p>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B4.png" alt="H3K36me3 Antibody for ChIP-seq assay" caption="false" width="700" height="107" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B5.png" alt="H3K36me3 Antibody validation in ChIP-seq " caption="false" width="700" height="114" /></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig1.png" alt="H3K36me3 Antibody for ChIP" caption="false" width="432" height="324" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. ChIP results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 µl per ChIP experiment was analysed. IgG (5 µg/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). 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="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig3.png" alt="H3K36me3 Antibody ELISA validation" caption="false" width="432" height="348" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig4.png" alt="H3K36me3 Antibody validated in Dot Blot" caption="false" width="278" height="186" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 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">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig5.png" alt="H3K36me3 Antibody validated in Western Blot" caption="false" width="278" height="354" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me3 </strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. </small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>5 - 10 µl/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:100 - 1:500</td>
<td>Fig 3</td>
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<tr>
<td>Dot Blotting</td>
<td>1:100,000</td>
<td>Fig 4</td>
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<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.</small></p>',
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$meta_title = 'H3K36me3 Antibody - ChIP-seq Grade (C15310058) | Diagenode'
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'id' => '2066',
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'name' => 'H3K36me3 Antibody - ChIP-seq Grade',
'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the trimethylated lysine 36 (H3K36me3), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2.png" alt="H3K36me3 Antibody ChIP Grade" caption="false" width="432" height="453" /></p>
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<p><small><strong>Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 µl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 µg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with 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 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes </small></p>
</div>
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<div class="row">
<div class="small-12 columns">
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B2.png" alt="H3K36me3 Antibody ChIP-seq Grade" caption="false" width="700" height="128" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B3.png" alt="H3K36me3 Antibody for ChIP-seq" caption="false" width="700" height="107" /></p>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B4.png" alt="H3K36me3 Antibody for ChIP-seq assay" caption="false" width="700" height="107" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B5.png" alt="H3K36me3 Antibody validation in ChIP-seq " caption="false" width="700" height="114" /></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig1.png" alt="H3K36me3 Antibody for ChIP" caption="false" width="432" height="324" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. ChIP results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 µl per ChIP experiment was analysed. IgG (5 µg/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). 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>
</div>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig3.png" alt="H3K36me3 Antibody ELISA validation" caption="false" width="432" height="348" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig4.png" alt="H3K36me3 Antibody validated in Dot Blot" caption="false" width="278" height="186" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 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">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig5.png" alt="H3K36me3 Antibody validated in Western Blot" caption="false" width="278" height="354" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me3 </strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. </small></p>
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<th>References</th>
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<tbody>
<tr>
<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>5 - 10 µl/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:100 - 1:500</td>
<td>Fig 3</td>
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<tr>
<td>Dot Blotting</td>
<td>1:100,000</td>
<td>Fig 4</td>
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<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.</small></p>',
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). 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. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<div class="row">
<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p></p>
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<p></p>
<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'description' => '<p>The interplay between methylation and demethylation of histone lysine residues is an essential component of gene expression regulation and there is considerable interest in elucidating the roles of proteins involved. Here we report that histone demethylase KDM4A/JMJD2A, which is involved in the regulation of cell proliferation and is overexpressed in some cancers, interacts with RNA Polymerase I, associates with active ribosomal RNA genes and is required for serum-induced activation of rDNA transcription. We propose that KDM4A controls the initial stages of transition from 'poised', non-transcribed rDNA chromatin into its active form. We show that PI3K, a major signalling transducer central for cell proliferation and survival, controls cellular localization of KDM4A and consequently its association with ribosomal DNA through the SGK1 downstream kinase. We propose that the interplay between PI3K/SGK1 signalling cascade and KDM4A constitutes a mechanism by which cells adapt ribosome biogenesis level to the availability of growth factors and nutrients.</p>',
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'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the trimethylated lysine 36 (H3K36me3), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2.png" alt="H3K36me3 Antibody ChIP Grade" caption="false" width="432" height="453" /></p>
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<p><small><strong>Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 µl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 µg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with 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 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes </small></p>
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<div class="small-12 columns">
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B2.png" alt="H3K36me3 Antibody ChIP-seq Grade" caption="false" width="700" height="128" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B3.png" alt="H3K36me3 Antibody for ChIP-seq" caption="false" width="700" height="107" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B4.png" alt="H3K36me3 Antibody for ChIP-seq assay" caption="false" width="700" height="107" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B5.png" alt="H3K36me3 Antibody validation in ChIP-seq " caption="false" width="700" height="114" /></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig1.png" alt="H3K36me3 Antibody for ChIP" caption="false" width="432" height="324" /></p>
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<p><small><strong>Figure 2. ChIP results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 µl per ChIP experiment was analysed. IgG (5 µg/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). 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><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig3.png" alt="H3K36me3 Antibody ELISA validation" caption="false" width="432" height="348" /></p>
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<p><small><strong>Figure 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700. </small></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me3 </strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. </small></p>
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'description' => '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. Trimethylation of histone H3K36 is preferentially present at active genes.',
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>5 - 10 µl/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:100 - 1:500</td>
<td>Fig 3</td>
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<td>Dot Blotting</td>
<td>1:100,000</td>
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<td>Western Blotting</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.</small></p>',
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<p><small><strong>Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 µl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 µg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with 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 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes </small></p>
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<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B3.png" alt="H3K36me3 Antibody for ChIP-seq" caption="false" width="700" height="107" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B4.png" alt="H3K36me3 Antibody for ChIP-seq assay" caption="false" width="700" height="107" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B5.png" alt="H3K36me3 Antibody validation in ChIP-seq " caption="false" width="700" height="114" /></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig1.png" alt="H3K36me3 Antibody for ChIP" caption="false" width="432" height="324" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 2. ChIP results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 µl per ChIP experiment was analysed. IgG (5 µg/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). 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>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig3.png" alt="H3K36me3 Antibody ELISA validation" caption="false" width="432" height="348" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700. </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="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig4.png" alt="H3K36me3 Antibody validated in Dot Blot" caption="false" width="278" height="186" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig5.png" alt="H3K36me3 Antibody validated in Western Blot" caption="false" width="278" height="354" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me3 </strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. </small></p>
</div>
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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. Trimethylation of histone H3K36 is preferentially present at active genes.</p>',
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'meta_title' => 'H3K36me3 Antibody - ChIP-seq Grade (C15310058) | Diagenode',
'meta_keywords' => '',
'meta_description' => 'H3K36me3 (Histone H3 trimethylated at lysine 36) Polyclonal Antibody validated in ChIP-seq, ChIP-grade, ELISA, DB and WB. Batch-specific data available on the website. ',
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'description' => '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. Trimethylation of histone H3K36 is preferentially present at active genes.',
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'lot' => 'A114-001 ',
'concentration' => 'Not determined',
'reactivity' => 'Human, mouse: positive. Other species: not tested.',
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<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>5 - 10 µl/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:100 - 1:500</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:100,000</td>
<td>Fig 4</td>
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<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
</tr>
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<p></p>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.</small></p>',
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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). 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. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
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<li>100% satisfaction guarantee</li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
<div class="small-12 medium-6 large-6 columns">
<p></p>
<p></p>
<p></p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the trimethylated lysine 36 (H3K36me3), using a KLH-conjugated synthetic peptide.</span></p>',
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2.png" alt="H3K36me3 Antibody ChIP Grade" caption="false" width="432" height="453" /></p>
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<p><small><strong>Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 µl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 µg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with 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 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes </small></p>
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<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B2.png" alt="H3K36me3 Antibody ChIP-seq Grade" caption="false" width="700" height="128" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B3.png" alt="H3K36me3 Antibody for ChIP-seq" caption="false" width="700" height="107" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B4.png" alt="H3K36me3 Antibody for ChIP-seq assay" caption="false" width="700" height="107" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B5.png" alt="H3K36me3 Antibody validation in ChIP-seq " caption="false" width="700" height="114" /></p>
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<p><small><strong>Figure 2. ChIP results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 µl per ChIP experiment was analysed. IgG (5 µg/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). 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 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700. </small></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me3 </strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. </small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>5 - 10 µl/ChIP</td>
<td>Fig 1, 2</td>
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<td>1:100 - 1:500</td>
<td>Fig 3</td>
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<td>Dot Blotting</td>
<td>1:100,000</td>
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<td>Western Blotting</td>
<td>1:1,000</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.</small></p>',
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<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2.png" alt="H3K36me3 Antibody ChIP Grade" caption="false" width="432" height="453" /></p>
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<p><small><strong>Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 µl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 µg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with 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 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes </small></p>
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<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B2.png" alt="H3K36me3 Antibody ChIP-seq Grade" caption="false" width="700" height="128" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B3.png" alt="H3K36me3 Antibody for ChIP-seq" caption="false" width="700" height="107" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B4.png" alt="H3K36me3 Antibody for ChIP-seq assay" caption="false" width="700" height="107" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B5.png" alt="H3K36me3 Antibody validation in ChIP-seq " caption="false" width="700" height="114" /></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig1.png" alt="H3K36me3 Antibody for ChIP" caption="false" width="432" height="324" /></p>
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<p><small><strong>Figure 2. ChIP results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 µl per ChIP experiment was analysed. IgG (5 µg/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). 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><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig3.png" alt="H3K36me3 Antibody ELISA validation" caption="false" width="432" height="348" /></p>
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<p><small><strong>Figure 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig4.png" alt="H3K36me3 Antibody validated in Dot Blot" caption="false" width="278" height="186" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 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">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig5.png" alt="H3K36me3 Antibody validated in Western Blot" caption="false" width="278" height="354" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me3 </strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. </small></p>
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<td>Fig 1, 2</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.</small></p>',
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). 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. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'description' => '<p><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'description' => 'Differentiation of female murine ES cells triggers silencing of one X chromosome through X-chromosome inactivation (XCI). Immunofluorescence studies showed that soon after Xist RNA coating the inactive X (Xi) undergoes many heterochromatic changes, including the acquisition of H3K27me3. However, the mechanisms that lead to the establishment of heterochromatin remain unclear. We first analyze chromatin changes by ChIP-chip, as well as RNA expression, around the X-inactivation center (Xic) in female and male ES cells, and their day 4 and 10 differentiated derivatives. A dynamic epigenetic landscape is observed within the Xic locus. Tsix repression is accompanied by deposition of H3K27me3 at its promoter during differentiation of both female and male cells. However, only in female cells does an active epigenetic landscape emerge at the Xist locus, concomitant with high Xist expression. Several regions within and around the Xic show unsuspected chromatin changes, and we define a series of unusual loci containing highly enriched H3K27me3. Genome-wide ChIP-seq analyses show a female-specific quantitative increase of H3K27me3 across the X chromosome as XCI proceeds in differentiating female ES cells. Using female ES cells with nonrandom XCI and polymorphic X chromosomes, we demonstrate that this increase is specific to the Xi by allele-specific SNP mapping of the ChIP-seq tags. H3K27me3 becomes evenly associated with the Xi in a chromosome-wide fashion. A selective and robust increase of H3K27me3 and concomitant decrease in H3K4me3 is observed over active genes. This indicates that deposition of H3K27me3 during XCI is tightly associated with the act of silencing of individual genes across the Xi.',
'date' => '2009-08-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/19581487',
'doi' => '',
'modified' => '2015-07-24 15:38:57',
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<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2.png" alt="H3K36me3 Antibody ChIP Grade" caption="false" width="432" height="453" /></p>
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<p><small><strong>Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 µl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 µg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with 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 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes </small></p>
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<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B2.png" alt="H3K36me3 Antibody ChIP-seq Grade" caption="false" width="700" height="128" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B3.png" alt="H3K36me3 Antibody for ChIP-seq" caption="false" width="700" height="107" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B4.png" alt="H3K36me3 Antibody for ChIP-seq assay" caption="false" width="700" height="107" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B5.png" alt="H3K36me3 Antibody validation in ChIP-seq " caption="false" width="700" height="114" /></p>
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<p><small><strong>Figure 2. ChIP results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 µl per ChIP experiment was analysed. IgG (5 µg/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). 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><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig3.png" alt="H3K36me3 Antibody ELISA validation" caption="false" width="432" height="348" /></p>
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<p><small><strong>Figure 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700. </small></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig5.png" alt="H3K36me3 Antibody validated in Western Blot" caption="false" width="278" height="354" /></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me3 </strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. </small></p>
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'classification' => 'Classic',
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<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
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<tbody>
<tr>
<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>5 - 10 µl/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:100 - 1:500</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:100,000</td>
<td>Fig 4</td>
</tr>
<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
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<p></p>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.</small></p>',
'storage_conditions' => 'Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.',
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'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the trimethylated lysine 36 (H3K36me3), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
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<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2.png" alt="H3K36me3 Antibody ChIP Grade" caption="false" width="432" height="453" /></p>
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<p><small><strong>Figure 1. ChIP-seq results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP was performed with 5 µl of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) on sheared chromatin from 1 million HeLaS3 cells using the “Auto Histone ChIP-seq” kit (cat. No. AB-Auto02-A100) on the IP-Star automated system. IgG (2 µg/IP) was used as a negative IP control. The IP’d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat (figure 2A). The IP’d DNA was subsequently analysed with 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 2B shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes </small></p>
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<div class="small-12 columns">
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B2.png" alt="H3K36me3 Antibody ChIP-seq Grade" caption="false" width="700" height="128" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B3.png" alt="H3K36me3 Antibody for ChIP-seq" caption="false" width="700" height="107" /></p>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B4.png" alt="H3K36me3 Antibody for ChIP-seq assay" caption="false" width="700" height="107" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig2B5.png" alt="H3K36me3 Antibody validation in ChIP-seq " caption="false" width="700" height="114" /></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig1.png" alt="H3K36me3 Antibody for ChIP" caption="false" width="432" height="324" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. ChIP results obtained with the Diagenode antibody directed against H3K36me3 </strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (cat. No. kch-oneDIP-060), using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 µl per ChIP experiment was analysed. IgG (5 µg/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). 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-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig3.png" alt="H3K36me3 Antibody ELISA validation" caption="false" width="432" height="348" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Determination of the titer </strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K36me3 (cat. No. CS-058-100). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:12,700. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15310058-Fig4.png" alt="H3K36me3 Antibody validated in Dot Blot" caption="false" width="278" height="186" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K36me3 </strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K36me3 (cat. No. CS-058-050) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. Figure 4 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="small-4 columns">
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K36me3 </strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K36me3 (cat. No. CS-058-100) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. </small></p>
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'description' => '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. Trimethylation of histone H3K36 is preferentially present at active genes.',
'clonality' => '',
'isotype' => '',
'lot' => 'A114-001 ',
'concentration' => 'Not determined',
'reactivity' => 'Human, mouse: positive. Other species: not tested.',
'type' => 'Polyclonal',
'purity' => 'Whole antiserum from rabbit containing 0.05% azide.',
'classification' => 'Classic',
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<thead>
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<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>5 - 10 µl/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:100 - 1:500</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:100,000</td>
<td>Fig 4</td>
</tr>
<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
</tr>
</tbody>
</table>
<p></p>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 µl per IP.</small></p>',
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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). 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. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
</div>
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<p>Diagenode’s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'description' => '<p><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
</div>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
<div class="small-12 medium-6 large-6 columns">
<p></p>
<p></p>
<p></p>
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<p></p>
<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'name' => 'The histone demethylase JMJD2A/KDM4A links ribosomal RNA transcription to nutrients and growth factors availability',
'authors' => 'Salifou K, Ray S, Verrier L, Aguirrebengoa M, Trouche D, Panov KI, Vandromme M',
'description' => '<p>The interplay between methylation and demethylation of histone lysine residues is an essential component of gene expression regulation and there is considerable interest in elucidating the roles of proteins involved. Here we report that histone demethylase KDM4A/JMJD2A, which is involved in the regulation of cell proliferation and is overexpressed in some cancers, interacts with RNA Polymerase I, associates with active ribosomal RNA genes and is required for serum-induced activation of rDNA transcription. We propose that KDM4A controls the initial stages of transition from 'poised', non-transcribed rDNA chromatin into its active form. We show that PI3K, a major signalling transducer central for cell proliferation and survival, controls cellular localization of KDM4A and consequently its association with ribosomal DNA through the SGK1 downstream kinase. We propose that the interplay between PI3K/SGK1 signalling cascade and KDM4A constitutes a mechanism by which cells adapt ribosome biogenesis level to the availability of growth factors and nutrients.</p>',
'date' => '2016-01-05',
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'description' => 'Mouse embryonic stem (ES) cells grown in serum exhibit greater heterogeneity in morphology and expression of pluripotency factors than ES cells cultured in defined medium with inhibitors of two kinases (Mek and GSK3), a condition known as "2i" postulated to establish a naive ground state. We show that the transcriptome and epigenome profiles of serum- and 2i-grown ES cells are distinct. 2i-treated cells exhibit lower expression of lineage-affiliated genes, reduced prevalence at promoters of the repressive histone modification H3K27me3, and fewer bivalent domains, which are thought to mark genes poised for either up- or downregulation. Nonetheless, serum- and 2i-grown ES cells have similar differentiation potential. Precocious transcription of developmental genes in 2i is restrained by RNA polymerase II promoter-proximal pausing. These findings suggest that transcriptional potentiation and a permissive chromatin context characterize the ground state and that exit from it may not require a metastable intermediate or multilineage priming.',
'date' => '2012-04-27',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/22541430',
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'description' => 'Differentiation of female murine ES cells triggers silencing of one X chromosome through X-chromosome inactivation (XCI). Immunofluorescence studies showed that soon after Xist RNA coating the inactive X (Xi) undergoes many heterochromatic changes, including the acquisition of H3K27me3. However, the mechanisms that lead to the establishment of heterochromatin remain unclear. We first analyze chromatin changes by ChIP-chip, as well as RNA expression, around the X-inactivation center (Xic) in female and male ES cells, and their day 4 and 10 differentiated derivatives. A dynamic epigenetic landscape is observed within the Xic locus. Tsix repression is accompanied by deposition of H3K27me3 at its promoter during differentiation of both female and male cells. However, only in female cells does an active epigenetic landscape emerge at the Xist locus, concomitant with high Xist expression. Several regions within and around the Xic show unsuspected chromatin changes, and we define a series of unusual loci containing highly enriched H3K27me3. Genome-wide ChIP-seq analyses show a female-specific quantitative increase of H3K27me3 across the X chromosome as XCI proceeds in differentiating female ES cells. Using female ES cells with nonrandom XCI and polymorphic X chromosomes, we demonstrate that this increase is specific to the Xi by allele-specific SNP mapping of the ChIP-seq tags. H3K27me3 becomes evenly associated with the Xi in a chromosome-wide fashion. A selective and robust increase of H3K27me3 and concomitant decrease in H3K4me3 is observed over active genes. This indicates that deposition of H3K27me3 during XCI is tightly associated with the act of silencing of individual genes across the Xi.',
'date' => '2009-08-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/19581487',
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[main] - APP/webroot/index.php, line 118
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