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<td>0.5-1 μg/ChIP</td>
<td>Fig 1, 2</td>
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<td><span mce-data-marked="1">ELISA</span></td>
<td>1:1,000</td>
<td>Fig 3</td>
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<td><span>1:1,000</span></td>
<td>Fig 4</td>
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<td><span>1:500</span></td>
<td>Fig 5</td>
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<td><span>1:200</span></td>
<td>Fig 6</td>
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<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-elisa.png" alt="H4K12ac Antibody ELISA validation" caption="false" width="400" height="302" /></div>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
<div class="small-8 columns">
<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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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<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
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<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<th>References</th>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>0.5-1 μg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td><span mce-data-marked="1">ELISA</span></td>
<td>1:1,000</td>
<td>Fig 3</td>
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<tr>
<td>Dot <span>Blotting</span></td>
<td><span>1:1,000</span></td>
<td>Fig 4</td>
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<tr>
<td><span>Western Blotting</span></td>
<td><span>1:500</span></td>
<td>Fig 5</td>
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<tr>
<td>IF</td>
<td><span>1:200</span></td>
<td>Fig 6</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H4 containing the acetylated lysines 12 (H4K12ac)</strong>, using a KLH-conjugated synthetic peptide. </p>',
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<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
</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 antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
</div>
<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-elisa.png" alt="H4K12ac Antibody ELISA validation" caption="false" width="400" height="302" /></div>
<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
<div class="small-8 columns">
<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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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<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
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<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </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 provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
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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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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p>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>
<ul>
<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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<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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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p></p>
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<p></p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'name' => 'Mutant FUS induces chromatin reorganization in the hippocampus andalters memory processes.',
'authors' => 'Tzeplaeff L. et al.',
'description' => '<p>Cytoplasmic mislocalization of the nuclear Fused in Sarcoma (FUS) protein is associated to amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Cytoplasmic FUS accumulation is recapitulated in the frontal cortex and spinal cord of heterozygous Fus mice. Yet, the mechanisms linking FUS mislocalization to hippocampal function and memory formation are still not characterized. Herein, we show that in these mice, the hippocampus paradoxically displays nuclear FUS accumulation. Multi-omic analyses showed that FUS binds to a set of genes characterized by the presence of an ETS/ELK-binding motifs, and involved in RNA metabolism, transcription, ribosome/mitochondria and chromatin organization. Importantly, hippocampal nuclei showed a decompaction of the neuronal chromatin at highly expressed genes and an inappropriate transcriptomic response was observed after spatial training of Fus mice. Furthermore, these mice lacked precision in a hippocampal-dependent spatial memory task and displayed decreased dendritic spine density. These studies shows that mutated FUS affects epigenetic regulation of the chromatin landscape in hippocampal neurons, which could participate in FTD/ALS pathogenic events. These data call for further investigation in the neurological phenotype of FUS-related diseases and open therapeutic strategies towards epigenetic drugs.</p>',
'date' => '2023-06-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/37327984',
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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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<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
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<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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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<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
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<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-elisa.png" alt="H4K12ac Antibody ELISA validation" caption="false" width="400" height="302" /></div>
<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
</div>
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<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
<div class="small-8 columns">
<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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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<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </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 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>
<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><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>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'meta_description' => 'Polyclonal and Monoclonal Antibodies against Histones and their modifications validated for many applications, including Chromatin Immunoprecipitation (ChIP) and ChIP-Sequencing (ChIP-seq)',
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'description' => '<h1><strong>Validated epigenetics antibodies</strong> – care for a sample?<br /> </h1>
<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>
<ul>
<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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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p></p>
<p></p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
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<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>0.5-1 μg/ChIP</td>
<td>Fig 1, 2</td>
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<td><span mce-data-marked="1">ELISA</span></td>
<td>1:1,000</td>
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<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
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<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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'description' => 'Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of histone H4 is associated with active genes.',
'clonality' => '',
'isotype' => '',
'lot' => 'A2439P',
'concentration' => '0.6 μg/μl',
'reactivity' => 'Human, mouse, wide range expected',
'type' => 'Polyclonal',
'purity' => 'Affinity purified',
'classification' => 'Classic',
'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>0.5-1 μg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td><span mce-data-marked="1">ELISA</span></td>
<td>1:1,000</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Dot <span>Blotting</span></td>
<td><span>1:1,000</span></td>
<td>Fig 4</td>
</tr>
<tr>
<td><span>Western Blotting</span></td>
<td><span>1:500</span></td>
<td>Fig 5</td>
</tr>
<tr>
<td>IF</td>
<td><span>1:200</span></td>
<td>Fig 6</td>
</tr>
</tbody>
</table>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
'storage_conditions' => 'Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.',
'storage_buffer' => '',
'precautions' => 'This product is for research use only. Not for use in diagnostic or therapeutic procedures.',
'uniprot_acc' => '',
'slug' => 'h4k12ac-polyclonal-antibody',
'meta_keywords' => '',
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'modified' => '2019-09-09 16:06:59',
'created' => '2016-12-12 16:39:41',
'select_label' => '499 - H4K12ac polyclonal antibody (A2439P - 0.6 μg/μl - Human, mouse, wide range expected - Affinity purified - Rabbit)'
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'Master' => array(
'id' => '2863',
'antibody_id' => '499',
'name' => 'H4K12ac Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H4 containing the acetylated lysines 12 (H4K12ac)</strong>, using a KLH-conjugated synthetic peptide. </p>',
'label1' => 'Validation data',
'info1' => '<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
<div class="small-6 columns">
<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
</div>
<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-elisa.png" alt="H4K12ac Antibody ELISA validation" caption="false" width="400" height="302" /></div>
<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
<div class="small-8 columns">
<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>
</div>',
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'info2' => '<p>Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of histone H4 is associated with active genes.</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 provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<p><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p>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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<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><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><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
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<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<th>References</th>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>0.5-1 μg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td><span mce-data-marked="1">ELISA</span></td>
<td>1:1,000</td>
<td>Fig 3</td>
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<td><span>1:500</span></td>
<td>Fig 5</td>
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<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
<div class="small-8 columns">
<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>
</div>',
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'info2' => '<p>Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of histone H4 is associated with active genes.</p>',
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'meta_title' => 'H4K12ac Antibody - ChIP-seq Grade (C15410331) | Diagenode',
'meta_keywords' => '',
'meta_description' => 'H4K12ac (Histone H4 acetylated at lysine 12) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR WB, ELISA, DB and IF. Batch-specific data available on the website. ',
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'created' => '2016-12-14 10:49:26',
'locale' => 'eng'
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'name' => 'H4K12ac polyclonal antibody',
'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. Acetylation of histone H4 is associated with active genes.',
'clonality' => '',
'isotype' => '',
'lot' => 'A2439P',
'concentration' => '0.6 μg/μl',
'reactivity' => 'Human, mouse, wide range expected',
'type' => 'Polyclonal',
'purity' => 'Affinity purified',
'classification' => 'Classic',
'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>0.5-1 μg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td><span mce-data-marked="1">ELISA</span></td>
<td>1:1,000</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Dot <span>Blotting</span></td>
<td><span>1:1,000</span></td>
<td>Fig 4</td>
</tr>
<tr>
<td><span>Western Blotting</span></td>
<td><span>1:500</span></td>
<td>Fig 5</td>
</tr>
<tr>
<td>IF</td>
<td><span>1:200</span></td>
<td>Fig 6</td>
</tr>
</tbody>
</table>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
'storage_conditions' => 'Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.',
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'precautions' => 'This product is for research use only. Not for use in diagnostic or therapeutic procedures.',
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'modified' => '2019-09-09 16:06:59',
'created' => '2016-12-12 16:39:41',
'select_label' => '499 - H4K12ac polyclonal antibody (A2439P - 0.6 μg/μl - Human, mouse, wide range expected - Affinity purified - Rabbit)'
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'name' => 'H4K12ac Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H4 containing the acetylated lysines 12 (H4K12ac)</strong>, using a KLH-conjugated synthetic peptide. </p>',
'label1' => 'Validation data',
'info1' => '<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
<div class="small-6 columns">
<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
</div>
<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-elisa.png" alt="H4K12ac Antibody ELISA validation" caption="false" width="400" height="302" /></div>
<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
<div class="small-8 columns">
<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
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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><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><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Cost-effective (requires less antibody per reaction)</li>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p>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><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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'id' => '2863',
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'name' => 'H4K12ac Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H4 containing the acetylated lysines 12 (H4K12ac)</strong>, using a KLH-conjugated synthetic peptide. </p>',
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<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<div class="small-12 columns">
<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-elisa.png" alt="H4K12ac Antibody ELISA validation" caption="false" width="400" height="302" /></div>
<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
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<div class="small-8 columns">
<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
<div class="small-8 columns">
<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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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<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
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<div class="small-12 columns">
<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone <strong>H4 containing the acetylated lysines 12 (H4K12ac)</strong>, using a KLH-conjugated synthetic peptide. </p>',
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<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-chip.png" alt="H4K12ac Antibody ChIP Grade" caption="false" width="400" height="303" /></div>
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<p><small> <strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H4K12ac (Cat. No. C15410331) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for promoter of the active GAPDH and EIF4A2 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="row">
<div class="small-12 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-a.png" alt="H4K12ac Antibody ChIP-seq Grade" caption="false" width="700" height="87" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-b.png" alt="H4K12ac Antibody for ChIP-seq " caption="false" width="700" height="191" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-c.png" alt="H4K12ac Antibody for ChIP-seq assay" caption="false" width="700" height="191" /></p>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/c15410331-chipseq-d.png" alt="H4K12ac Antibody validated in ChIP-seq " caption="false" width="700" height="161" /></p>
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<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H4K12ac</strong><br /> ChIP was performed with 0.5 μg of the Diagenode antibody against H4K12ac (Cat. No. C15410331) on sheared chromatin from 1,000,000 HeLa cells using the “iDeal ChIP-seq” kit as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq 2000. 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 peak distribution along the complete sequence and a 2 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D). </small></p>
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<div class="row">
<div class="small-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-elisa.png" alt="H4K12ac Antibody ELISA validation" caption="false" width="400" height="302" /></div>
<div class="small-6 columns">
<p><small> <strong>Figure 3. Determination of the antibody titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H4K12ac (Cat. No. C15410331) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 3), the titer of the antibody was estimated to be 1:14,300. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-dotblot.png" alt="H4K12ac Antibody Dot Blot validation" caption="false" width="298" height="308" /></p>
</div>
<div class="small-8 columns">
<p><small> <strong>Figure 4. Cross reactivity tests using the Diagenode antibody directed against H4K12ac</strong><br /> To test the cross reactivity of the Diagenode antibody against H4K12ac (Cat. No. C15410331), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 4 shows the antibody is specific for the K12 acetylation with some slight cross reaction with K8ac. </small></p>
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<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/c15410331-wb.png" alt="H4K12ac Antibody validated in Western Blot" caption="false" width="297" height="222" /></div>
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<p><small> <strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H4K12ac</strong><br /> Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode antibody against H4K12ac (Cat. No. C15410331). The antibody was diluted 1:500 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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<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410331-if.png" alt="H4K12ac Antibody validated in Immunofluorescence" caption="false" width="400" height="97" /></p>
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<div class="row">
<div class="small-12 columns">
<p><small> <strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H4K12ac</strong><br /> HeLa cells were stained with the Diagenode antibody against H4K12ac (Cat. No. C15410331) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K12ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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'description' => '<p>Polyclonal antibody raised in rabbit against the region of histone H4 containing the acetylated lysines 12 (H4K12ac), using a KLH-conjugated synthetic peptide.</p>',
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'description' => '<p>Cytoplasmic mislocalization of the nuclear Fused in Sarcoma (FUS) protein is associated to amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Cytoplasmic FUS accumulation is recapitulated in the frontal cortex and spinal cord of heterozygous Fus mice. Yet, the mechanisms linking FUS mislocalization to hippocampal function and memory formation are still not characterized. Herein, we show that in these mice, the hippocampus paradoxically displays nuclear FUS accumulation. Multi-omic analyses showed that FUS binds to a set of genes characterized by the presence of an ETS/ELK-binding motifs, and involved in RNA metabolism, transcription, ribosome/mitochondria and chromatin organization. Importantly, hippocampal nuclei showed a decompaction of the neuronal chromatin at highly expressed genes and an inappropriate transcriptomic response was observed after spatial training of Fus mice. Furthermore, these mice lacked precision in a hippocampal-dependent spatial memory task and displayed decreased dendritic spine density. These studies shows that mutated FUS affects epigenetic regulation of the chromatin landscape in hippocampal neurons, which could participate in FTD/ALS pathogenic events. These data call for further investigation in the neurological phenotype of FUS-related diseases and open therapeutic strategies towards epigenetic drugs.</p>',
'date' => '2023-06-01',
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