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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3</strong><br />ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns">A.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2A-ChIP-seq.jpg" alt="H3K27me3 Antibody ChIP-seq Grade" /></center><br /> B.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2B-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq" />
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H3K27me3 (cat. No. C15210017) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 signal distribution along the complete sequence of chromosome 1 and a zoomin to a 3 Mb region (fig 2A and B) and in two genomic regions surrounding the MYT1 and TSH2B positive control genes (fig 2C and D, respectively).</p>
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<p><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K27me3 (cat. No. C15210017), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. 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 2 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27me3 (cat. No. C15210017). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3</strong><br />ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<p><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K27me3 (cat. No. C15210017), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. 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 2 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27me3 (cat. No. C15210017). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.</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><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-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15210018-chip.jpg" alt="H3K27me3 Antibody ChIP Grade" /></center></div>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3</strong><br />ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns">A.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2A-ChIP-seq.jpg" alt="H3K27me3 Antibody ChIP-seq Grade" /></center><br /> B.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2B-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq" />
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</center><br /> C.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2C-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq assay" /></center><br /> D.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2D-ChIP-seq.jpg" alt="H3K27me3 Antibody validated in ChIP-seq" /></center></div>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H3K27me3 (cat. No. C15210017) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 signal distribution along the complete sequence of chromosome 1 and a zoomin to a 3 Mb region (fig 2A and B) and in two genomic regions surrounding the MYT1 and TSH2B positive control genes (fig 2C and D, respectively).</p>
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<p><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K27me3 (cat. No. C15210017), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. 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 2 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27me3 (cat. No. C15210017). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15210018-chip.jpg" alt="H3K27me3 Antibody ChIP Grade" /></center></div>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3</strong><br />ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns">A.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2A-ChIP-seq.jpg" alt="H3K27me3 Antibody ChIP-seq Grade" /></center><br /> B.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2B-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq" />
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</center><br /> C.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2C-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq assay" /></center><br /> D.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2D-ChIP-seq.jpg" alt="H3K27me3 Antibody validated in ChIP-seq" /></center></div>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H3K27me3 (cat. No. C15210017) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 signal distribution along the complete sequence of chromosome 1 and a zoomin to a 3 Mb region (fig 2A and B) and in two genomic regions surrounding the MYT1 and TSH2B positive control genes (fig 2C and D, respectively).</p>
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<p><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K27me3 (cat. No. C15210017), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. 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 2 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27me3 (cat. No. C15210017). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.</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’s highly validated antibodies:</span></p>
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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>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>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><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>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-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15210018-chip.jpg" alt="H3K27me3 Antibody ChIP Grade" /></center></div>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3</strong><br />ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns">A.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2A-ChIP-seq.jpg" alt="H3K27me3 Antibody ChIP-seq Grade" /></center><br /> B.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2B-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq" />
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</center><br /> C.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2C-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq assay" /></center><br /> D.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2D-ChIP-seq.jpg" alt="H3K27me3 Antibody validated in ChIP-seq" /></center></div>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H3K27me3 (cat. No. C15210017) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 signal distribution along the complete sequence of chromosome 1 and a zoomin to a 3 Mb region (fig 2A and B) and in two genomic regions surrounding the MYT1 and TSH2B positive control genes (fig 2C and D, respectively).</p>
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<p><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K27me3 (cat. No. C15210017), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. 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 2 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27me3 (cat. No. C15210017). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.</p>
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'label1' => 'Validation data',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15210018-chip.jpg" alt="H3K27me3 Antibody ChIP Grade" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3</strong><br />ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="row">
<div class="small-12 columns">A.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2A-ChIP-seq.jpg" alt="H3K27me3 Antibody ChIP-seq Grade" /></center><br /> B.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2B-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq" />
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</center><br /> C.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2C-ChIP-seq.jpg" alt="H3K27me3 Antibody for ChIP-seq assay" /></center><br /> D.<center><img src="https://www.diagenode.com/img/product/antibodies/C15210017-fig2D-ChIP-seq.jpg" alt="H3K27me3 Antibody validated in ChIP-seq" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K27me3</strong><br />ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 0.5 µg of the Diagenode antibody against H3K27me3 (cat. No. C15210017) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 signal distribution along the complete sequence of chromosome 1 and a zoomin to a 3 Mb region (fig 2A and B) and in two genomic regions surrounding the MYT1 and TSH2B positive control genes (fig 2C and D, respectively).</p>
<p></p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15210018-db.jpg" alt="H3K27me3 Antibody validated in Dot Blot" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K27me3 (cat. No. C15210017), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. 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 2 shows a high specificity of the antibody for the modification of interest.</p>
</div>
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<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/c15210018-wb.jpg" alt="H3K27me3 Antibody validated in Western blot" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27me3 (cat. No. C15210017). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.</p>
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'description' => 'Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called "histone code". Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Trimethylation of histone H3K27 is associated with inactive genomic regions.',
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>0.5 - 1 µg per IP</td>
<td>Fig 1, 2</td>
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<td>Dot Blotting</td>
<td>1:20,000</td>
<td>Fig 3</td>
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<td>Western Blotting</td>
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<td>Fig 4</td>
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<p><small>*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5 - 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’s highly validated antibodies:</span></p>
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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><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
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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><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3</strong><br />ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<p><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />To test the cross reactivity of the Diagenode antibody against H3K27me3 (cat. No. C15210017), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K27. 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 2 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K27me3</strong><br />Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K27me3 (cat. No. C15210017). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K27me3</strong><br />ChIP was performed with the Diagenode antibody against H3K27me3 (cat. No. C15210017) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the TSH2B and MYT1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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'description' => 'Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called "histone code". Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Trimethylation of histone H3K27 is associated with inactive genomic regions.',
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>0.5 - 1 µg per IP</td>
<td>Fig 1, 2</td>
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<p><small>*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5 - 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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<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><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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×
