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'meta_description' => 'H3K27me3S28p (Histone H3 trimethylated at lysine 27 and phosphorylated at serine 28) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, ELISA, DB and WB. ',
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'description' => '<p><span>Polyclonal antibody raised in rabbit against against histone H3, trimethylated at lysine 27 and phosphorylated at serine 28 (H3K27me3S28p), 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/C15410091_ChIP.jpg" alt="H3K27me3S28p Antibody ChIP Grade" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million 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 the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-A.jpg" alt="H3K27me3S28p Antibody ChIP-seq Grade" caption="false" width="432" height="127" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-B.jpg" alt="H3K27me3S28p Antibody for ChIP-seq" caption="false" width="432" height="103" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-C.jpg" alt="H3K27me3S28p Antibody for ChIP-seq assay" caption="false" width="432" height="86" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-D.jpg" alt="H3K27me3S28p Antibody validated in ChIP-seq" caption="false" width="432" height="103" /></p>
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<div class="small-6 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP was performed on sheared chromatin from 1.5 million colcemid treated HeLaS3 cells using 2 μg of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) 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 in genomic regions surrounding the TSH2B and MYT1 positive control genes (fig 2A and B, indicated by an arrow), and of chromosome 3 and 20 (figure 2C and D). </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ELISA.jpg" alt="H3K27me3S28p Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. 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 H3K27me3S28p (Cat. No. C15410091). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:4,300. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_DotBlot.jpg" alt="H3K27me3S28p Antibody validated in Dot Blot" caption="false" width="278" height="169" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Cross reactivity test of the Diagenode antibody directed against H3K27me3S28p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 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/C15410091_WB-A.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="221" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_WB-B.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="171" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Western blot analysis using the Diagenode antibody directed against H3K27me3S28p</strong><br /> A. Western blot was performed on histone extracts (15 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) diluted 1:100 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. B. Cell cycle experiment using the Diagenode antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. </small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<td>ELISA</td>
<td>1:200</td>
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<td>1:2,000</td>
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<td>1:1,000</td>
<td>Fig 5</td>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIP.jpg" alt="H3K27me3S28p Antibody ChIP Grade" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million 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 the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes. </small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-A.jpg" alt="H3K27me3S28p Antibody ChIP-seq Grade" caption="false" width="432" height="127" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-B.jpg" alt="H3K27me3S28p Antibody for ChIP-seq" caption="false" width="432" height="103" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-C.jpg" alt="H3K27me3S28p Antibody for ChIP-seq assay" caption="false" width="432" height="86" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-D.jpg" alt="H3K27me3S28p Antibody validated in ChIP-seq" caption="false" width="432" height="103" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP was performed on sheared chromatin from 1.5 million colcemid treated HeLaS3 cells using 2 μg of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) 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 in genomic regions surrounding the TSH2B and MYT1 positive control genes (fig 2A and B, indicated by an arrow), and of chromosome 3 and 20 (figure 2C and D). </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ELISA.jpg" alt="H3K27me3S28p Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. 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 H3K27me3S28p (Cat. No. C15410091). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:4,300. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_DotBlot.jpg" alt="H3K27me3S28p Antibody validated in Dot Blot" caption="false" width="278" height="169" /></p>
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<p><small><strong> Figure 4. Cross reactivity test of the Diagenode antibody directed against H3K27me3S28p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 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/C15410091_WB-A.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="221" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_WB-B.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="171" /></p>
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<p><small><strong> Figure 5. Western blot analysis using the Diagenode antibody directed against H3K27me3S28p</strong><br /> A. Western blot was performed on histone extracts (15 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) diluted 1:100 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. B. Cell cycle experiment using the Diagenode antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. </small></p>
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<td>Fig 4</td>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<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>
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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>Batch-specific data is available on the website</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>
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<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 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>
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<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 H3K27me3S28p</strong><br /> ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million 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 the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-A.jpg" alt="H3K27me3S28p Antibody ChIP-seq Grade" caption="false" width="432" height="127" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP was performed on sheared chromatin from 1.5 million colcemid treated HeLaS3 cells using 2 μg of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) 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 in genomic regions surrounding the TSH2B and MYT1 positive control genes (fig 2A and B, indicated by an arrow), and of chromosome 3 and 20 (figure 2C and D). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ELISA.jpg" alt="H3K27me3S28p Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 3. 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 H3K27me3S28p (Cat. No. C15410091). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:4,300. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_DotBlot.jpg" alt="H3K27me3S28p Antibody validated in Dot Blot" caption="false" width="278" height="169" /></p>
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<p><small><strong> Figure 4. Cross reactivity test of the Diagenode antibody directed against H3K27me3S28p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 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/C15410091_WB-A.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="221" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_WB-B.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="171" /></p>
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<p><small><strong> Figure 5. Western blot analysis using the Diagenode antibody directed against H3K27me3S28p</strong><br /> A. Western blot was performed on histone extracts (15 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) diluted 1:100 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. B. Cell cycle experiment using the Diagenode antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. </small></p>
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<td>Fig 1, 2</td>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIP.jpg" alt="H3K27me3S28p Antibody ChIP Grade" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million 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 the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-A.jpg" alt="H3K27me3S28p Antibody ChIP-seq Grade" caption="false" width="432" height="127" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-B.jpg" alt="H3K27me3S28p Antibody for ChIP-seq" caption="false" width="432" height="103" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-C.jpg" alt="H3K27me3S28p Antibody for ChIP-seq assay" caption="false" width="432" height="86" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP was performed on sheared chromatin from 1.5 million colcemid treated HeLaS3 cells using 2 μg of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) 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 in genomic regions surrounding the TSH2B and MYT1 positive control genes (fig 2A and B, indicated by an arrow), and of chromosome 3 and 20 (figure 2C and D). </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ELISA.jpg" alt="H3K27me3S28p Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 3. 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 H3K27me3S28p (Cat. No. C15410091). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:4,300. </small></p>
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<p><small><strong> Figure 4. Cross reactivity test of the Diagenode antibody directed against H3K27me3S28p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 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/C15410091_WB-A.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="221" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_WB-B.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="171" /></p>
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<p><small><strong> Figure 5. Western blot analysis using the Diagenode antibody directed against H3K27me3S28p</strong><br /> A. Western blot was performed on histone extracts (15 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) diluted 1:100 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. B. Cell cycle experiment using the Diagenode antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<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>
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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>Batch-specific data is available on the website</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>
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<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>
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<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;"> 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><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIP.jpg" alt="H3K27me3S28p Antibody ChIP Grade" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million 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 the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-A.jpg" alt="H3K27me3S28p Antibody ChIP-seq Grade" caption="false" width="432" height="127" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-B.jpg" alt="H3K27me3S28p Antibody for ChIP-seq" caption="false" width="432" height="103" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-C.jpg" alt="H3K27me3S28p Antibody for ChIP-seq assay" caption="false" width="432" height="86" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-D.jpg" alt="H3K27me3S28p Antibody validated in ChIP-seq" caption="false" width="432" height="103" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP was performed on sheared chromatin from 1.5 million colcemid treated HeLaS3 cells using 2 μg of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) 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 in genomic regions surrounding the TSH2B and MYT1 positive control genes (fig 2A and B, indicated by an arrow), and of chromosome 3 and 20 (figure 2C and D). </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ELISA.jpg" alt="H3K27me3S28p Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. 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 H3K27me3S28p (Cat. No. C15410091). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:4,300. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_DotBlot.jpg" alt="H3K27me3S28p Antibody validated in Dot Blot" caption="false" width="278" height="169" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Cross reactivity test of the Diagenode antibody directed against H3K27me3S28p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 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/C15410091_WB-A.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="221" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_WB-B.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="171" /></p>
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<p><small><strong> Figure 5. Western blot analysis using the Diagenode antibody directed against H3K27me3S28p</strong><br /> A. Western blot was performed on histone extracts (15 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) diluted 1:100 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. B. Cell cycle experiment using the Diagenode antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. </small></p>
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<th>Suggested dilution</th>
<th>References</th>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:200</td>
<td>Fig 3</td>
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<tr>
<td>Dot Blotting</td>
<td>1:2,000</td>
<td>Fig 4</td>
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<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIP.jpg" alt="H3K27me3S28p Antibody ChIP Grade" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million 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 the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes. </small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-A.jpg" alt="H3K27me3S28p Antibody ChIP-seq Grade" caption="false" width="432" height="127" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-B.jpg" alt="H3K27me3S28p Antibody for ChIP-seq" caption="false" width="432" height="103" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-C.jpg" alt="H3K27me3S28p Antibody for ChIP-seq assay" caption="false" width="432" height="86" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-D.jpg" alt="H3K27me3S28p Antibody validated in ChIP-seq" caption="false" width="432" height="103" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP was performed on sheared chromatin from 1.5 million colcemid treated HeLaS3 cells using 2 μg of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) 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 in genomic regions surrounding the TSH2B and MYT1 positive control genes (fig 2A and B, indicated by an arrow), and of chromosome 3 and 20 (figure 2C and D). </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ELISA.jpg" alt="H3K27me3S28p Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. 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 H3K27me3S28p (Cat. No. C15410091). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:4,300. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_DotBlot.jpg" alt="H3K27me3S28p Antibody validated in Dot Blot" caption="false" width="278" height="169" /></p>
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<p><small><strong> Figure 4. Cross reactivity test of the Diagenode antibody directed against H3K27me3S28p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 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/C15410091_WB-A.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="221" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_WB-B.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="171" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. Western blot analysis using the Diagenode antibody directed against H3K27me3S28p</strong><br /> A. Western blot was performed on histone extracts (15 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) diluted 1:100 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. B. Cell cycle experiment using the Diagenode antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. </small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:200</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:2,000</td>
<td>Fig 4</td>
</tr>
<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
</tr>
</tbody>
</table>
<p><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>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></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>
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<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>
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<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>
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<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><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIP.jpg" alt="H3K27me3S28p Antibody ChIP Grade" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million 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 the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-A.jpg" alt="H3K27me3S28p Antibody ChIP-seq Grade" caption="false" width="432" height="127" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-B.jpg" alt="H3K27me3S28p Antibody for ChIP-seq" caption="false" width="432" height="103" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP was performed on sheared chromatin from 1.5 million colcemid treated HeLaS3 cells using 2 μg of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) 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 in genomic regions surrounding the TSH2B and MYT1 positive control genes (fig 2A and B, indicated by an arrow), and of chromosome 3 and 20 (figure 2C and D). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ELISA.jpg" alt="H3K27me3S28p Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 3. 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 H3K27me3S28p (Cat. No. C15410091). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:4,300. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_DotBlot.jpg" alt="H3K27me3S28p Antibody validated in Dot Blot" caption="false" width="278" height="169" /></p>
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<p><small><strong> Figure 4. Cross reactivity test of the Diagenode antibody directed against H3K27me3S28p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 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/C15410091_WB-A.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="221" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_WB-B.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="171" /></p>
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<p><small><strong> Figure 5. Western blot analysis using the Diagenode antibody directed against H3K27me3S28p</strong><br /> A. Western blot was performed on histone extracts (15 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) diluted 1:100 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. B. Cell cycle experiment using the Diagenode antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. </small></p>
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<th>Suggested dilution</th>
<th>References</th>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:200</td>
<td>Fig 3</td>
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<td>Fig 4</td>
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<td>Western Blotting</td>
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<td>Fig 5</td>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIP.jpg" alt="H3K27me3S28p Antibody ChIP Grade" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP assays were performed using human HeLa cells treated with colcemid, the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million 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 the promoter of the active gene GAPDH as a negative control, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes. </small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-A.jpg" alt="H3K27me3S28p Antibody ChIP-seq Grade" caption="false" width="432" height="127" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-B.jpg" alt="H3K27me3S28p Antibody for ChIP-seq" caption="false" width="432" height="103" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-C.jpg" alt="H3K27me3S28p Antibody for ChIP-seq assay" caption="false" width="432" height="86" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ChIPSeq-D.jpg" alt="H3K27me3S28p Antibody validated in ChIP-seq" caption="false" width="432" height="103" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3S28p</strong><br /> ChIP was performed on sheared chromatin from 1.5 million colcemid treated HeLaS3 cells using 2 μg of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) 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 in genomic regions surrounding the TSH2B and MYT1 positive control genes (fig 2A and B, indicated by an arrow), and of chromosome 3 and 20 (figure 2C and D). </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_ELISA.jpg" alt="H3K27me3S28p Antibody ELISA validation" caption="false" width="278" height="210" /></p>
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<p><small><strong> Figure 3. 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 H3K27me3S28p (Cat. No. C15410091). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:4,300. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_DotBlot.jpg" alt="H3K27me3S28p Antibody validated in Dot Blot" caption="false" width="278" height="169" /></p>
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<p><small><strong> Figure 4. Cross reactivity test of the Diagenode antibody directed against H3K27me3S28p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000. Figure 3 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/C15410091_WB-A.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="221" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410091_WB-B.jpg" alt="H3K27me3S28p Antibody validated in Western Blot" caption="false" width="278" height="171" /></p>
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<p><small><strong> Figure 5. Western blot analysis using the Diagenode antibody directed against H3K27me3S28p</strong><br /> A. Western blot was performed on histone extracts (15 μg, lane 1) from HeLa cells, and on 1 μg of recombinant histone H3 (lane 2) using the Diagenode antibody against H3K27me3S28p (Cat. No. C15410091) diluted 1:100 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. B. Cell cycle experiment using the Diagenode antibody against H3K27me3S28p. WB was performed on cells blocked in different stages of the cell cycle (1=asynchronous population, 2=G1/Sa, 3=G1/Sb, 4=S phase, 5=G2/M, 6=M, 7=G1). The picture shows a nice peak of H3K27me3S28p expression during mitosis. </small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<td>ELISA</td>
<td>1:200</td>
<td>Fig 3</td>
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<tr>
<td>Dot Blotting</td>
<td>1:2,000</td>
<td>Fig 4</td>
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<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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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>
<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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<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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)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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