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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</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><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
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<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><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>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>
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-chip.png" alt="H3K14ac Antibody ChIP Grade" caption="false" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
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<div class="extra-spaced"></div>
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<div class="small-12 columns">A.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-a.jpg" alt="H3K14ac Antibody ChIP-seq Grade" caption="false" width="893" height="106" /><br /> B.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-b.jpg" alt="H3K14ac Antibody for ChIP-seq " caption="false" width="893" height="268" /><br /> C.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-c.jpg" alt="H3K14ac Antibody for ChIP-seq assay" caption="false" width="893" height="166" /><br /> D.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-d.jpg" alt="H3K14ac Antibody validated in ChIP-seq " caption="false" width="893" height="193" /></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-dotblot.png" alt="H3K14ac Antibody validated in Dot Blot" caption="false" width="201" height="261" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-wb.png" alt="H3K14ac Antibody validated in Western Blot" caption="false" width="201" height="319" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-if.png" alt="H3K14ac Antibody validated in Immunofluorescence" caption="false" width="198" height="198" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</small></p>
</div>
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'application_table' => '<table>
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<th>Suggested dilution</th>
<th>References</th>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1 μg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>Dot blot</td>
<td>1:10,000</td>
<td>Fig 3</td>
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<tr>
<td>Western Blotting</td>
<td>1:2,000</td>
<td>Fig 4</td>
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<td>Immunofluorescence</td>
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<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><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
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<div class="small-12 columns">A.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-a.jpg" alt="H3K14ac Antibody ChIP-seq Grade" caption="false" width="893" height="106" /><br /> B.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-b.jpg" alt="H3K14ac Antibody for ChIP-seq " caption="false" width="893" height="268" /><br /> C.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-c.jpg" alt="H3K14ac Antibody for ChIP-seq assay" caption="false" width="893" height="166" /><br /> D.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-d.jpg" alt="H3K14ac Antibody validated in ChIP-seq " caption="false" width="893" height="193" /></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).</small></p>
</div>
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<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-dotblot.png" alt="H3K14ac Antibody validated in Dot Blot" caption="false" width="201" height="261" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-wb.png" alt="H3K14ac Antibody validated in Western Blot" caption="false" width="201" height="319" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-if.png" alt="H3K14ac Antibody validated in Immunofluorescence" caption="false" width="198" height="198" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</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>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<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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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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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><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><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 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/c15210005-wb.png" alt="H3K14ac Antibody validated in Western Blot" caption="false" width="201" height="319" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-if.png" alt="H3K14ac Antibody validated in Immunofluorescence" caption="false" width="198" height="198" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</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>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<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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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><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>
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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>
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> - Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
<li><strong>Strict quality standards</strong> with rigorous QC and validation</li>
<li><strong>Classified</strong> based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</small></p>
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<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><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>
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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 has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> - Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
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<li><strong>Classified</strong> based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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'description' => '<p>Monoclonal antibody raised in rabbit against histone <strong>H3 acetylated at Lys14 (H3K14ac),</strong> using a KLH-conjugated synthetic peptide.</p>',
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-chip.png" alt="H3K14ac Antibody ChIP Grade" caption="false" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="small-12 columns">A.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-a.jpg" alt="H3K14ac Antibody ChIP-seq Grade" caption="false" width="893" height="106" /><br /> B.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-b.jpg" alt="H3K14ac Antibody for ChIP-seq " caption="false" width="893" height="268" /><br /> C.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-c.jpg" alt="H3K14ac Antibody for ChIP-seq assay" caption="false" width="893" height="166" /><br /> D.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-d.jpg" alt="H3K14ac Antibody validated in ChIP-seq " caption="false" width="893" height="193" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-dotblot.png" alt="H3K14ac Antibody validated in Dot Blot" caption="false" width="201" height="261" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 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/c15210005-wb.png" alt="H3K14ac Antibody validated in Western Blot" caption="false" width="201" height="319" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-if.png" alt="H3K14ac Antibody validated in Immunofluorescence" caption="false" width="198" height="198" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</small></p>
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'name' => 'H3K14ac Antibody',
'description' => '<p>Monoclonal antibody raised in rabbit against histone <strong>H3 acetylated at Lys14 (H3K14ac),</strong> using a KLH-conjugated synthetic peptide.</p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-chip.png" alt="H3K14ac Antibody ChIP Grade" caption="false" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP assays were performed using HeLa cells, the Diagenode antibody against H3K14ac (cat. No. C15210005) 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 consisting of 0.5, 1, 2 and 5 µg of antibody per ChIP experiment was analyzed. IgG (1 µg/IP) was used as a negative IP control. Quantitative PCR was performed with optimized primers for the promoters of the EIF2S3 and GAPDH genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="small-12 columns">A.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-a.jpg" alt="H3K14ac Antibody ChIP-seq Grade" caption="false" width="893" height="106" /><br /> B.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-b.jpg" alt="H3K14ac Antibody for ChIP-seq " caption="false" width="893" height="268" /><br /> C.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-c.jpg" alt="H3K14ac Antibody for ChIP-seq assay" caption="false" width="893" height="166" /><br /> D.<img src="https://www.diagenode.com/img/product/antibodies/c15210005-chipseq-d.jpg" alt="H3K14ac Antibody validated in ChIP-seq " caption="false" width="893" height="193" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K14ac</strong><br />ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 μg of the Diagenode antibody against H3K14ac (Cat. No. C15210005) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 1 Mb region of human chromosome 3 (fig 2A and B), and in two genomic regions surrounding the EIF2S3 and GAPDH positive control genes (fig 2C and D).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-dotblot.png" alt="H3K14ac Antibody validated in Dot Blot" caption="false" width="201" height="261" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) with peptides containing different modifications or unmodified sequences of histone H3. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:10,000. Figure 2 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/c15210005-wb.png" alt="H3K14ac Antibody validated in Western Blot" caption="false" width="201" height="319" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />Histone extracts from HeLa cells treated with butyrate (lane 2) or untreated control cells (lane 1) were analysed by Western blot using the Diagenode monoclonal antibody against H3K14ac (Cat. No. C15210005) diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15210005-if.png" alt="H3K14ac Antibody validated in Immunofluorescence" caption="false" width="198" height="198" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K14ac</strong><br />HeLa cells were stained with the Diagenode antibody against H3K14ac (Cat. No. C15210005 red) diluted 1:500. Actin filaments were stained with fluorescein phalladoin (green).</small></p>
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'info2' => '<p>Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the socalled “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of Lys14 is associated with gene activation.</p>',
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