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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_chip.png" /></center></div>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-A.png" alt="H3K4me3 Antibody ChIP-seq Grade" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-B.png" alt="H3K4me3 Antibody for ChIP-seq" /></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_ELISA.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_IF.png" /></center></div>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<td>2 μg/ChIP</td>
<td>Fig 1,2</td>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_chip.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-A.png" alt="H3K4me3 Antibody ChIP-seq Grade" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-B.png" alt="H3K4me3 Antibody for ChIP-seq" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-C.png" alt="H3K4me3 Antibody for ChIP-seq assay" /></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152-cuttag-a.png" /></p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
</div>
</div>
<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_ELISA.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_WB.png" /></center></div>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_IF.png" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><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><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>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> - Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
<li><strong>Strict quality standards</strong> with rigorous QC and validation</li>
<li><strong>Classified</strong> based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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'description' => '<p>BACKGROUND: Interactions of chromatin with the nuclear lamina via lamina-associated domains (LADs) confer structural stability to the genome. The dynamics of positioning of LADs during differentiation, and how LADs impinge on developmental gene expression, remains, however, elusive. RESULTS: We examined changes in the association of lamin B1 with the genome in the first 72 h of differentiation of adipose stem cells into adipocytes. We demonstrate a repositioning of entire stand-alone LADs and of LAD edges as a prominent nuclear structural feature of early adipogenesis. Whereas adipogenic genes are released from LADs, LADs sequester downregulated or repressed genes irrelevant for the adipose lineage. However, LAD repositioning only partly concurs with gene expression changes. Differentially expressed genes in LADs, including LADs conserved throughout differentiation, reside in local euchromatic and lamin-depleted sub-domains. In these sub-domains, pre-differentiation histone modification profiles correlate with the LAD versus inter-LAD outcome of these genes during adipogenic commitment. Lastly, we link differentially expressed genes in LADs to short-range enhancers which overall co-partition with these genes in LADs versus inter-LADs during differentiation. CONCLUSIONS: We conclude that LADs are predictable structural features of adipose nuclear architecture that restrain non-adipogenic genes in a repressive environment.</p>',
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_chip.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-A.png" alt="H3K4me3 Antibody ChIP-seq Grade" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-B.png" alt="H3K4me3 Antibody for ChIP-seq" /></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152-cuttag-a.png" /></p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_ELISA.png" /></center></div>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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'name' => 'H3K4me3 Antibody',
'description' => '<p><span>Monoclonal antibody raised in mouse against histone <strong>H3, trimethylated at lysine 4</strong> (<strong>H3K4me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_chip.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-A.png" alt="H3K4me3 Antibody ChIP-seq Grade" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-B.png" alt="H3K4me3 Antibody for ChIP-seq" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-C.png" alt="H3K4me3 Antibody for ChIP-seq assay" /></p>
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<p>D.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-D.png" alt="H3K4me3 Antibody validated in ChIP-seq " /></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152-cuttag-a.png" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/C15200152-cuttag-b.png" /></p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_ELISA.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_WB.png" /></center></div>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_IF.png" /></center></div>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</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><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<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>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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'name' => 'H3K4me3 Antibody',
'description' => '<p><span>Monoclonal antibody raised in mouse against histone <strong>H3, trimethylated at lysine 4</strong> (<strong>H3K4me3</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_chip.png" /></center></div>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-A.png" alt="H3K4me3 Antibody ChIP-seq Grade" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-B.png" alt="H3K4me3 Antibody for ChIP-seq" /></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152-cuttag-a.png" /></p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_WB.png" /></center></div>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p>C.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-C.png" alt="H3K4me3 Antibody for ChIP-seq assay" /></p>
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<p>D.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-D.png" alt="H3K4me3 Antibody validated in ChIP-seq " /></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p>A.<img src="https://www.diagenode.com/img/product/antibodies/C15200152-cuttag-a.png" /></p>
<p>B.<img src="https://www.diagenode.com/img/product/antibodies/C15200152-cuttag-b.png" /></p>
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<div class="small-12 columns">
<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<div class="row">
<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_ELISA.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_WB.png" /></center></div>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_IF.png" /></center></div>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_chip.png" /></center></div>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p>B.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-B.png" alt="H3K4me3 Antibody for ChIP-seq" /></p>
<p>C.<img src="https://www.diagenode.com/img/product/antibodies/C15200152_chipseq-C.png" alt="H3K4me3 Antibody for ChIP-seq assay" /></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_ELISA.png" /></center></div>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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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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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<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><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> - Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
<li><strong>Strict quality standards</strong> with rigorous QC and validation</li>
<li><strong>Classified</strong> based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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'name' => 'H3K4me3 Antibody',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_chip.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_chip.png" /></center></div>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>',
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<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>'
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_ELISA.png" /></center></div>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
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<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<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><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> - Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong></p>
<p>ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) and optimized PCR primer pairs for qPCR. ChIP was performed with the “Auto Histone ChIP-seq” kit (Cat. No. C01010022), using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the constitutively expressed GAPDH and c-fos genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene 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). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 2 μg of the Diagenode antibody against H3K4me3 (Cat. No. C15200152) as described above. 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. Figure 2 shows the peak distribution along two 5 Mb regions of chromosome 3 and 5 (figure 2A and B, respectively) and in two 100 kb regions surrounding the GAPDH and c-fos positive control genes (figure 2C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.</p>
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<p><strong>Figure 3. Cut&Tag results obtained with the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> CUT&TAG (Kaya-Okur, H.S., Nat Commun 10, 1930, 2019) was performed on 50,000 K562 cells using 1 µg of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152) and the Diagenode pA-Tn5 transposase (C01070001). The libraries were subsequently analysed on an Illumina NextSeq 500 sequencer (2x75 paired-end reads) according to the manufacturer's instructions. The tags were aligned to the human genome (hg19) using the BWA algorithm. Figure 3 shows the peak distribution along the complete sequence and a 1.5 Mb zoomin of chromosome 1 (figure 3A and B, respectively).</p>
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<p><strong>Figure 4. Cross reactivity of the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against H3K4me3 (cat. No. C15200152). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure 4 shows a high specificity of the antibody for the modification of interest.</p>
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<p><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> Histone extracts (15 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K4me3 (Cat. No. C15200152) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</p>
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<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15200152_IF.png" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against H3K4me3</strong><br /> HeLa cells were stained with the Diagenode antibody against H3K4me3 (Cat. No. C15200152) 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 labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</p>
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>',
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<p>Read more:</p>
<p><a href="https://www.diagenode.com/en/categories/cutandtag">Products for CUT&Tag assay</a></p>
<p><a href="https://www.diagenode.com/en/pages/cut-and-tag">Performance of Diagenode's antibodies in CUT&Tag</a></p>'
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'date' => '2021-02-01',
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View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118
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