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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-chip.jpg" alt="H3K56ac Antibody ChIP Grade" caption="false" width="278" height="202" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<td style="height: 42px;">ChIP/ChIP-seq <sup>*</sup></td>
<td style="height: 42px;">5 μg/ChIP</td>
<td style="height: 42px;">Fig 1, 2</td>
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<td style="height: 37px;">Cut&Tag</td>
<td style="height: 37px;">1 μg</td>
<td style="height: 37px;">Fig 3</td>
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<td style="height: 37px;">ELISA</td>
<td style="height: 37px;">1:500</td>
<td style="height: 37px;">Fig 4</td>
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<td style="height: 37px;">Dot Blotting</td>
<td style="height: 37px;">1:20,000</td>
<td style="height: 37px;">Fig 5</td>
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<td style="height: 37px;">Western Blotting</td>
<td style="height: 37px;">1:200</td>
<td style="height: 37px;">Fig 6</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 1-5 μg per IP.</small></p>',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-8 columns">
<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<td style="height: 42px;">ChIP/ChIP-seq <sup>*</sup></td>
<td style="height: 42px;">5 μg/ChIP</td>
<td style="height: 42px;">Fig 1, 2</td>
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<tr style="height: 37px;">
<td style="height: 37px;">Cut&Tag</td>
<td style="height: 37px;">1 μg</td>
<td style="height: 37px;">Fig 3</td>
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<tr style="height: 37px;">
<td style="height: 37px;">ELISA</td>
<td style="height: 37px;">1:500</td>
<td style="height: 37px;">Fig 4</td>
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<tr style="height: 37px;">
<td style="height: 37px;">Dot Blotting</td>
<td style="height: 37px;">1:20,000</td>
<td style="height: 37px;">Fig 5</td>
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<tr style="height: 37px;">
<td style="height: 37px;">Western Blotting</td>
<td style="height: 37px;">1:200</td>
<td style="height: 37px;">Fig 6</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 1-5 μg per IP.</small></p>',
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'name' => 'H3K56ac Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the acetylated lysine 56 (H3K56ac)</strong>, using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-chip.jpg" alt="H3K56ac Antibody ChIP Grade" caption="false" width="278" height="202" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-8 columns">
<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
</div>
<div class="small-4 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p></p>
<div class="row">
<div class="small-8 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
</div>
<br />
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<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>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></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>
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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>
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<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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<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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<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>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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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-chip.jpg" alt="H3K56ac Antibody ChIP Grade" caption="false" width="278" height="202" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-8 columns">
<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
</div>
<div class="small-4 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
</div>
</div>
<p></p>
<div class="row">
<div class="small-8 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
</div>
<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
</div>
<br />
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>',
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>'
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'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the acetylated lysine 56 (H3K56ac)</strong>, using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-chip.jpg" alt="H3K56ac Antibody ChIP Grade" caption="false" width="278" height="202" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-8 columns">
<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
</div>
<div class="small-4 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p></p>
<div class="row">
<div class="small-8 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<br />
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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'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.',
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<thead>
<tr style="height: 55px;">
<th style="height: 55px;">Applications</th>
<th style="height: 55px;">Suggested dilution</th>
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<td style="height: 42px;">ChIP/ChIP-seq <sup>*</sup></td>
<td style="height: 42px;">5 μg/ChIP</td>
<td style="height: 42px;">Fig 1, 2</td>
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<tr style="height: 37px;">
<td style="height: 37px;">Cut&Tag</td>
<td style="height: 37px;">1 μg</td>
<td style="height: 37px;">Fig 3</td>
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<td style="height: 37px;">ELISA</td>
<td style="height: 37px;">1:500</td>
<td style="height: 37px;">Fig 4</td>
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<td style="height: 37px;">Dot Blotting</td>
<td style="height: 37px;">1:20,000</td>
<td style="height: 37px;">Fig 5</td>
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<td style="height: 37px;">Western Blotting</td>
<td style="height: 37px;">1:200</td>
<td style="height: 37px;">Fig 6</td>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<td style="height: 42px;">ChIP/ChIP-seq <sup>*</sup></td>
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<td style="height: 37px;">Fig 4</td>
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<td style="height: 37px;">Dot Blotting</td>
<td style="height: 37px;">1:20,000</td>
<td style="height: 37px;">Fig 5</td>
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<td style="height: 37px;">Western Blotting</td>
<td style="height: 37px;">1:200</td>
<td style="height: 37px;">Fig 6</td>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<div class="row">
<div class="small-4 columns">
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></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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<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>
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<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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<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>
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<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><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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<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>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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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<div class="row">
<div class="small-8 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<br />
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<br />
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
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'description' => '<p>More than in any other immuoprecipitation assays, quality antibodies are critical tools in many epigenetics experiments. Since 10 years, Diagenode has developed the most stringent quality production available on the market for antibodies exclusively focused on epigenetic uses. All our antibodies have been qualified to work in epigenetic applications.</p>',
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'name' => 'Quantification and epigenetic evaluation of the residual pool of hepatitisB covalently closed circular DNA in long-term nucleoside analogue-treatedpatients.',
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'description' => '<p>Hepatitis B virus (HBV) covalently closed circular (ccc)DNA is the key genomic form responsible for viral persistence and virological relapse after treatment withdrawal. The assessment of residual intrahepatic cccDNA levels and activity after long-term nucleos(t)ide analogues therapy still represents a technical challenge. Quantitative (q)PCR, rolling circle amplification (RCA) and droplet digital (dd)PCR assays were used to quantify residual intrahepatic cccDNA in liver biopsies from 56 chronically HBV infected patients after 3 to 5 years of telbivudine treatment. Activity of residual cccDNA was evaluated by quantifying 3.5 kB HBV RNA (preC/pgRNA) and by assessing cccDNA-associated histone tails post-transcriptional modifications (PTMs) by micro-chromatin immunoprecipitation. Long-term telbivudine treatment resulted in serum HBV DNA suppression, with most of the patients reaching undetectable levels. Despite 38 out of 56 patients had undetectable cccDNA when assessed by qPCR, RCA and ddPCR assays detected cccDNA in all-but-one negative samples. Low preC/pgRNA level in telbivudine-treated samples was associated with enrichment for cccDNA histone PTMs related to repressed transcription. No difference in cccDNA levels was found according to serum viral markers evolution. This panel of cccDNA evaluation techniques should provide an added value for the new proof-of-concept clinical trials aiming at a functional cure of chronic hepatitis B.</p>',
'date' => '2020-12-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/33273565',
'doi' => '10.1038/s41598-020-78001-1',
'modified' => '2022-01-06 14:53:18',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<th style="height: 55px;">Applications</th>
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<td style="height: 42px;">ChIP/ChIP-seq <sup>*</sup></td>
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<td style="height: 37px;">Fig 5</td>
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<td style="height: 37px;">Fig 6</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 1-5 μg per IP.</small></p>',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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'meta_title' => 'H3K56ac Antibody - ChIP-seq Grade (C15410213) | Diagenode',
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'meta_description' => 'H3K56ac (Histone H3 acetylated at lysine 56) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, ELISA and WB. Batch-specific data available on the website. Sample size available.',
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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.',
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'isotype' => '',
'lot' => 'A2069P',
'concentration' => '0.8 µg/µl',
'reactivity' => 'Human',
'type' => 'Polyclonal, <strong>ChIP grade/ChIP-seq grade</strong>',
'purity' => 'Affinity purified',
'classification' => 'Classic',
'application_table' => '<table>
<thead>
<tr style="height: 55px;">
<th style="height: 55px;">Applications</th>
<th style="height: 55px;">Suggested dilution</th>
<th style="height: 55px;">References</th>
</tr>
</thead>
<tbody>
<tr style="height: 42px;">
<td style="height: 42px;">ChIP/ChIP-seq <sup>*</sup></td>
<td style="height: 42px;">5 μg/ChIP</td>
<td style="height: 42px;">Fig 1, 2</td>
</tr>
<tr style="height: 37px;">
<td style="height: 37px;">Cut&Tag</td>
<td style="height: 37px;">1 μg</td>
<td style="height: 37px;">Fig 3</td>
</tr>
<tr style="height: 37px;">
<td style="height: 37px;">ELISA</td>
<td style="height: 37px;">1:500</td>
<td style="height: 37px;">Fig 4</td>
</tr>
<tr style="height: 37px;">
<td style="height: 37px;">Dot Blotting</td>
<td style="height: 37px;">1:20,000</td>
<td style="height: 37px;">Fig 5</td>
</tr>
<tr style="height: 37px;">
<td style="height: 37px;">Western Blotting</td>
<td style="height: 37px;">1:200</td>
<td style="height: 37px;">Fig 6</td>
</tr>
</tbody>
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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 1-5 μg per IP.</small></p>',
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'name' => 'H3K56ac Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the acetylated lysine 56 (H3K56ac)</strong>, using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-chip.jpg" alt="H3K56ac Antibody ChIP Grade" caption="false" width="278" height="202" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-8 columns">
<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
</div>
<div class="small-4 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
</div>
</div>
<p></p>
<div class="row">
<div class="small-8 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
</div>
<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
</div>
<br />
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<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>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></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>
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<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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<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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'name' => 'H3K56ac Antibody (sample size)',
'description' => '<p><span>Polyclonal antibody raised in rabbit against the region of histone <strong>H3 containing the acetylated lysine 56 (H3K56ac)</strong>, using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-chip.jpg" alt="H3K56ac Antibody ChIP Grade" caption="false" width="278" height="202" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<br />
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
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<div class="small-4 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<br />
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>'
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p></p>
<div class="row">
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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</div>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<td style="height: 42px;">ChIP/ChIP-seq <sup>*</sup></td>
<td style="height: 42px;">5 μg/ChIP</td>
<td style="height: 42px;">Fig 1, 2</td>
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<td style="height: 37px;">Cut&Tag</td>
<td style="height: 37px;">1 μg</td>
<td style="height: 37px;">Fig 3</td>
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<td style="height: 37px;">ELISA</td>
<td style="height: 37px;">1:500</td>
<td style="height: 37px;">Fig 4</td>
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<td style="height: 37px;">Dot Blotting</td>
<td style="height: 37px;">1:20,000</td>
<td style="height: 37px;">Fig 5</td>
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<td style="height: 37px;">Western Blotting</td>
<td style="height: 37px;">1:200</td>
<td style="height: 37px;">Fig 6</td>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<td style="height: 42px;">ChIP/ChIP-seq <sup>*</sup></td>
<td style="height: 42px;">5 μg/ChIP</td>
<td style="height: 42px;">Fig 1, 2</td>
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<td style="height: 37px;">Cut&Tag</td>
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<td style="height: 37px;">Fig 3</td>
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<td style="height: 37px;">1:500</td>
<td style="height: 37px;">Fig 4</td>
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<td style="height: 37px;">Dot Blotting</td>
<td style="height: 37px;">1:20,000</td>
<td style="height: 37px;">Fig 5</td>
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<td style="height: 37px;">Western Blotting</td>
<td style="height: 37px;">1:200</td>
<td style="height: 37px;">Fig 6</td>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-a.jpg" alt="H3K56ac Antibody ChIP-seq Grade" caption="false" width="550" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-b.jpg" alt="H3K56ac Antibody for ChIP-seq" caption="false" width="550" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-c.jpg" alt="H3K56ac Antibody for ChIP-seq assay" caption="false" width="550" /></p>
<p>D.<img src="https://www.diagenode.com/img/product/antibodies/c15410213-chipseq-d.jpg" alt="H3K56ac Antibody validated in ChIP-seq" caption="false" width="550" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<div class="row">
<div class="small-8 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-cuttag.jpg" alt="H3K56ac Antibody Cut and tag" caption="false" width="550" /></p>
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<div class="small-4 columns">
<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
</div>
<br />
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-elisa.jpg" alt="H3K56ac Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-wb.jpg" alt="H3K56ac Antibody validated in Western Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<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>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></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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<p>Diagenode’s highly validated antibodies:</p>
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<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>
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<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><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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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP assays were performed using human K562 cells, the Diagenode antibody against H3K56ac (cat. No. C15410213) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 500,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for GAPDH and EIF4A2 promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> ChIP was performed on sheared chromatin from 500,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig 2A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.</small></p>
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<p><small><strong> Figure 3. Cut&Tag results obtained with the Diagenode antibody directed against H3K56ac</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode antibody against H3K56ac (cat. No. C15410213) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer’s instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution in 2 genomic regions surrounding the FOS gene on chromosome 14 and the CCT5 gene on chromosome 5 (figure 3A and B, respectively).</small></p>
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<p><small><strong> Figure 4. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K56ac (cat. No. 15410213) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 4), the titer of the antibody was estimated to be 1:36,500. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410213-db.jpg" alt="H3K56ac Antibody validated in Dot Blot " caption="false" width="278" height="341" /></p>
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<p><small><strong> Figure 5. Cross reactivity tests using the Diagenode antibody directed against H3K56ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H3K56ac (cat. No. 15410213), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 5 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 6.Western blot analysis using the Diagenode antibody directed against H3K56ac</strong><br /> Western blot was performed on whole cell extracts (40 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K56ac (cat. No. C15410213). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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'name' => 'CUT&Tag',
'description' => '<p>CUT＆Tagアッセイを成功させるための重要な要素の1つは使用される抗体の品質です。 特異性高い抗体は、目的のタンパク質のみをターゲットとした確実な結果を可能にします。 CUT＆Tagで検証済みの抗体のセレクションはこちらからご覧ください。</p>
<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>
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'name' => 'CUT&Tag',
'description' => '<p>The quality of antibody used in CUT&Tag is one of the crucial factors for assay success. The antibodies with confirmed high specificity will target only the protein of interest, enabling real results. Check out our selection of antibodies validated in CUT&Tag.</p>
<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>',
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'modified' => '2021-04-27 05:17:46',
'created' => '2020-08-20 10:13:47',
'locale' => 'eng'
)
$description = '<p>The quality of antibody used in CUT&Tag is one of the crucial factors for assay success. The antibodies with confirmed high specificity will target only the protein of interest, enabling real results. Check out our selection of antibodies validated in CUT&Tag.</p>
<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>'
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'name' => 'Epigenetic Antibodies Brochure',
'description' => '<p>More than in any other immuoprecipitation assays, quality antibodies are critical tools in many epigenetics experiments. Since 10 years, Diagenode has developed the most stringent quality production available on the market for antibodies exclusively focused on epigenetic uses. All our antibodies have been qualified to work in epigenetic applications.</p>',
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'name' => 'SDS C15410213 H3K56ac Antibody FR fr',
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'name' => 'Quantification and epigenetic evaluation of the residual pool of hepatitisB covalently closed circular DNA in long-term nucleoside analogue-treatedpatients.',
'authors' => 'Lebossé F. et al.',
'description' => '<p>Hepatitis B virus (HBV) covalently closed circular (ccc)DNA is the key genomic form responsible for viral persistence and virological relapse after treatment withdrawal. The assessment of residual intrahepatic cccDNA levels and activity after long-term nucleos(t)ide analogues therapy still represents a technical challenge. Quantitative (q)PCR, rolling circle amplification (RCA) and droplet digital (dd)PCR assays were used to quantify residual intrahepatic cccDNA in liver biopsies from 56 chronically HBV infected patients after 3 to 5 years of telbivudine treatment. Activity of residual cccDNA was evaluated by quantifying 3.5 kB HBV RNA (preC/pgRNA) and by assessing cccDNA-associated histone tails post-transcriptional modifications (PTMs) by micro-chromatin immunoprecipitation. Long-term telbivudine treatment resulted in serum HBV DNA suppression, with most of the patients reaching undetectable levels. Despite 38 out of 56 patients had undetectable cccDNA when assessed by qPCR, RCA and ddPCR assays detected cccDNA in all-but-one negative samples. Low preC/pgRNA level in telbivudine-treated samples was associated with enrichment for cccDNA histone PTMs related to repressed transcription. No difference in cccDNA levels was found according to serum viral markers evolution. This panel of cccDNA evaluation techniques should provide an added value for the new proof-of-concept clinical trials aiming at a functional cure of chronic hepatitis B.</p>',
'date' => '2020-12-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/33273565',
'doi' => '10.1038/s41598-020-78001-1',
'modified' => '2022-01-06 14:53:18',
'created' => '2021-12-06 15:53:19',
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$externalLink = ' <a href="https://www.ncbi.nlm.nih.gov/pubmed/33273565" target="_blank"><i class="fa fa-external-link"></i></a>'include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118
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