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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIP.jpg" alt="H3K4ac Antibody ChIP Grade" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-A.jpg" alt="H3K4ac Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-B.jpg" alt="H3K4ac Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-C.jpg" alt="H3K4ac Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-D.jpg" alt="H3K4ac Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<div class="row">
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ELISA.jpg" alt="H3K4ac Antibody ELISA validation" /></p>
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<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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-dotblot.jpg" alt="H3K4ac Antibody Dot Blot validation" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-WB.jpg" alt="H3K4ac Antibody validated in Western Blot" /></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-IF.jpg" alt="H3K4ac Antibody validated in Immunofluorescence " /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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>Fig 1, 2</td>
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<td>1:1,000</td>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIP.jpg" alt="H3K4ac Antibody ChIP Grade" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-A.jpg" alt="H3K4ac Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-B.jpg" alt="H3K4ac Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-C.jpg" alt="H3K4ac Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-D.jpg" alt="H3K4ac Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ELISA.jpg" alt="H3K4ac Antibody ELISA validation" /></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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-dotblot.jpg" alt="H3K4ac Antibody Dot Blot validation" /></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIP.jpg" alt="H3K4ac Antibody ChIP Grade" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-A.jpg" alt="H3K4ac Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-B.jpg" alt="H3K4ac Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-C.jpg" alt="H3K4ac Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-D.jpg" alt="H3K4ac Antibody validated in ChIP-seq" /></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 monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two 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">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ELISA.jpg" alt="H3K4ac Antibody ELISA validation" /></p>
</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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-dotblot.jpg" alt="H3K4ac Antibody Dot Blot validation" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-WB.jpg" alt="H3K4ac Antibody validated in Western Blot" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-IF.jpg" alt="H3K4ac Antibody validated in Immunofluorescence " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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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'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.</p>',
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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
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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>
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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><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two 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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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 H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two 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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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 H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two 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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-dotblot.jpg" alt="H3K4ac Antibody Dot Blot validation" /></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<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>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p>Learn more about: <a href="../applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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'meta_keywords' => ' Polyclonal antibody,H3K4ac,histone H3 acetylated ',
'meta_description' => 'H3K4ac (Histone H3 acetylated at lysine 4) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, ELISA, DB, IF and WB. Batch-specific data available on the website. Sample size available.',
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'name' => 'H3K4ac Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against histone <strong>H3 acetylated at lysine 4 (H3K4ac)</strong>, using a KLH-conjugated synthetic peptide. </p>',
'label1' => 'Validation data',
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIP.jpg" alt="H3K4ac Antibody ChIP Grade" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-A.jpg" alt="H3K4ac Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-B.jpg" alt="H3K4ac Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-C.jpg" alt="H3K4ac Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-D.jpg" alt="H3K4ac Antibody validated in ChIP-seq" /></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 monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ELISA.jpg" alt="H3K4ac Antibody ELISA validation" /></p>
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<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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-dotblot.jpg" alt="H3K4ac Antibody Dot Blot validation" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-WB.jpg" alt="H3K4ac Antibody validated in Western Blot" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-IF.jpg" alt="H3K4ac Antibody validated in Immunofluorescence " /></p>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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>Suggested dilution</th>
<th>References</th>
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<td>ChIP/ChIP-seq<sup>*</sup></td>
<td>2 µg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:1,000 - 1:10,000</td>
<td>Fig 3</td>
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<tr>
<td>Dot Blotting</td>
<td>1:5,000</td>
<td>Fig 4</td>
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<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
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<tr>
<td>Immunofluorescence</td>
<td>1:200</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.</small></p>',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two 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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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>
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<td>1:200</td>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-A.jpg" alt="H3K4ac Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-B.jpg" alt="H3K4ac Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-C.jpg" alt="H3K4ac Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-D.jpg" alt="H3K4ac Antibody validated in ChIP-seq" /></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two 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">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ELISA.jpg" alt="H3K4ac Antibody ELISA validation" /></p>
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<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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-dotblot.jpg" alt="H3K4ac Antibody Dot Blot validation" /></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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>
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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 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><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p><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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'description' => '<p>Breast cancer is the most frequently diagnosed malignancy in women worldwide. It is well established that the complexity of carcinogenesis involves profound epigenetic deregulations that contribute to the tumorigenesis process. Deregulated H3 and H4 acetylated histone marks are amongst those alterations. Sirtuin-1 (SIRT1) is a class-III histone deacetylase deeply involved in apoptosis, genomic stability, gene expression regulation and breast tumorigenesis. However, the underlying molecular mechanism by which SIRT1 regulates H3 and H4 acetylated marks, and consequently cancer-related gene expression in breast cancer, remains uncharacterized. In this study, we elucidated SIRT1 epigenetic role and analyzed the link between the latter and histones H3 and H4 epigenetic marks in all 5 molecular subtypes of breast cancer. Using a cohort of 135 human breast tumors and their matched normal tissues, as well as 5 human-derived cell lines, we identified H3k4ac as a new prime target of SIRT1 in breast cancer. We also uncovered an inverse correlation between SIRT1 and the 3 epigenetic marks H3k4ac, H3k9ac and H4k16ac expression patterns. We showed that SIRT1 modulates the acetylation patterns of histones H3 and H4 in breast cancer. Moreover, SIRT1 regulates its H3 acetylated targets in a subtype-specific manner. Furthermore, SIRT1 siRNA-mediated knockdown increases histone acetylation levels at 6 breast cancer-related gene promoters: <em>AR</em>, <em>BRCA1</em>, <em>ERS1</em>, <em>ERS2</em>, <em>EZH2</em> and <em>EP300</em>. In summary, this report characterizes for the first time the epigenetic behavior of SIRT1 in human breast carcinoma. These novel findings point to a potential use of SIRT1 as an epigenetic therapeutic target in breast cancer.</p>',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two 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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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>
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<td>Fig 1, 2</td>
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<tr>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two 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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-dotblot.jpg" alt="H3K4ac Antibody Dot Blot validation" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
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<div class="extra-spaced"></div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-WB.jpg" alt="H3K4ac Antibody validated in Western Blot" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-IF.jpg" alt="H3K4ac Antibody validated in Immunofluorescence " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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>',
'label2' => 'Target Description',
'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.</p>',
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'meta_title' => 'H3K4ac Antibody - ChIP-seq Grade (C15410322) | Diagenode',
'meta_keywords' => ' Polyclonal antibody,H3K4ac,histone H3 acetylated ',
'meta_description' => 'H3K4ac (Histone H3 acetylated at lysine 4) Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, ELISA, DB, IF and WB. Batch-specific data available on the website. Sample size available.',
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'created' => '2015-12-02 15:43:54',
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'name' => 'H3K4ac polyclonal antibody - Classic',
'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.',
'clonality' => '',
'isotype' => '',
'lot' => 'A2505P',
'concentration' => '1.1 μg/μl',
'reactivity' => 'Human',
'type' => 'Polyclonal',
'purity' => 'Affinity purified polyclonal antibody',
'classification' => '',
'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>2 µg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:1,000 - 1:10,000</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:5,000</td>
<td>Fig 4</td>
</tr>
<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 5</td>
</tr>
<tr>
<td>Immunofluorescence</td>
<td>1:200</td>
<td>Fig 6</td>
</tr>
</tbody>
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<p></p>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user.</small></p>',
'storage_conditions' => 'Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.',
'storage_buffer' => 'PBS containing 0.05% azide and 0.05% ProClin 300.',
'precautions' => 'This product is for research use only. Not for use in diagnostic or therapeutic procedures.',
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'name' => 'H3K4ac Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against histone <strong>H3 acetylated at lysine 4 (H3K4ac)</strong>, using a KLH-conjugated synthetic peptide. </p>',
'label1' => 'Validation data',
'info1' => '<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIP.jpg" alt="H3K4ac Antibody ChIP Grade" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K4ac</strong><br />ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K4ac (Cat. No. C15410322) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as positive controls, and for the HBB promoter and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-A.jpg" alt="H3K4ac Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-B.jpg" alt="H3K4ac Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-C.jpg" alt="H3K4ac Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ChIPseq-D.jpg" alt="H3K4ac Antibody validated in ChIP-seq" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against TAL1</strong><br />ChIP was performed with 1 µg of the Diagenode antibody against H3K4ac (Cat. No. C15410322) on sheared chromatin from 4 million HeLa cells 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 1.5 mb region of the X chromosome (figure 2A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes (figure 2C and D).</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-ELISA.jpg" alt="H3K4ac Antibody ELISA validation" /></p>
</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 H3K4ac (Cat. No. C15410322) 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 purified antibody was estimated to be 1:197,500.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-dotblot.jpg" alt="H3K4ac Antibody Dot Blot validation" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 4. Cross reactivity test using the Diagenode antibody directed against H3K4ac</strong> <br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K4ac (Cat. No. C15410322) with peptides containing other histone H3 and H4 modifications and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest.</small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
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<div class="extra-spaced"></div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-WB.jpg" alt="H3K4ac Antibody validated in Western Blot" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode antibody directed against H3K4ac</strong> <br /> Whole cell (25 μg, lane 1) and histone extracts (15 µg, lane 2) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K4ac (Cat. No. C15410322), diluted 1:1,000 in TBSTween containing 5% BSA. The marker (in kDa) is shown on the left, the position of the protein of interest is indicated on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410322-IF.jpg" alt="H3K4ac Antibody validated in Immunofluorescence " /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode antibody directed against H3K4ac</strong> <br /> HeLa cells were stained with the Diagenode antibody against H3K4ac (Cat. No. C15410322) 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 H3K4ac 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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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<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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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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'name' => 'SIRT1-dependent epigenetic regulation of H3 and H4 histone acetylation in human breast cancer',
'authors' => 'Khaldoun Rifaï et al.',
'description' => '<p>Breast cancer is the most frequently diagnosed malignancy in women worldwide. It is well established that the complexity of carcinogenesis involves profound epigenetic deregulations that contribute to the tumorigenesis process. Deregulated H3 and H4 acetylated histone marks are amongst those alterations. Sirtuin-1 (SIRT1) is a class-III histone deacetylase deeply involved in apoptosis, genomic stability, gene expression regulation and breast tumorigenesis. However, the underlying molecular mechanism by which SIRT1 regulates H3 and H4 acetylated marks, and consequently cancer-related gene expression in breast cancer, remains uncharacterized. In this study, we elucidated SIRT1 epigenetic role and analyzed the link between the latter and histones H3 and H4 epigenetic marks in all 5 molecular subtypes of breast cancer. Using a cohort of 135 human breast tumors and their matched normal tissues, as well as 5 human-derived cell lines, we identified H3k4ac as a new prime target of SIRT1 in breast cancer. We also uncovered an inverse correlation between SIRT1 and the 3 epigenetic marks H3k4ac, H3k9ac and H4k16ac expression patterns. We showed that SIRT1 modulates the acetylation patterns of histones H3 and H4 in breast cancer. Moreover, SIRT1 regulates its H3 acetylated targets in a subtype-specific manner. Furthermore, SIRT1 siRNA-mediated knockdown increases histone acetylation levels at 6 breast cancer-related gene promoters: <em>AR</em>, <em>BRCA1</em>, <em>ERS1</em>, <em>ERS2</em>, <em>EZH2</em> and <em>EP300</em>. In summary, this report characterizes for the first time the epigenetic behavior of SIRT1 in human breast carcinoma. These novel findings point to a potential use of SIRT1 as an epigenetic therapeutic target in breast cancer.</p>',
'date' => '2018-07-17',
'pmid' => 'http://www.oncotarget.com/index.php?journal=oncotarget&page=article&op=view&path[]=25771&path[]=80619',
'doi' => '',
'modified' => '2018-08-09 10:47:58',
'created' => '2018-07-26 12:02:12',
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