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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.</small></p>',
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP-b.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ELISA.jpg" alt="H3K27me1 Antibody validated in ELISA" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-DotBlot.jpg" alt="H3K27me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<div class="row">
<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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'description' => 'Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases.',
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<thead>
<tr>
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<th>References</th>
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<tbody>
<tr>
<td>ChIP <sup>*</sup></td>
<td>1 µg/ChIP</td>
<td>Fig 1</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:500</td>
<td>Fig 2</td>
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<tr>
<td>Dot Blotting</td>
<td>1:20,000</td>
<td>Fig 3</td>
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<tr>
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<td>1:1,000</td>
<td>Fig 4</td>
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<td>1:1,000</td>
<td>Fig 5</td>
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<p></p>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.</small></p>',
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP-b.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ELISA.jpg" alt="H3K27me1 Antibody validated in ELISA" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-DotBlot.jpg" alt="H3K27me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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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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<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>
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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>To date, it remains largely unclear to what extent chromatin machinery contributes to the susceptibility and progression of complex diseases. Here, we combine deep epigenome mapping with single-cell transcriptomics to mine for evidence of chromatin dysregulation in type 2 diabetes. We find two chromatin-state signatures that track β cell dysfunction in mice and humans: ectopic activation of bivalent Polycomb-silenced domains and loss of expression at an epigenomically unique class of lineage-defining genes. β cell-specific Polycomb (Eed/PRC2) loss of function in mice triggers diabetes-mimicking transcriptional signatures and highly penetrant, hyperglycemia-independent dedifferentiation, indicating that PRC2 dysregulation contributes to disease. The work provides novel resources for exploring β cell transcriptional regulation and identifies PRC2 as necessary for long-term maintenance of β cell identity. Importantly, the data suggest a two-hit (chromatin and hyperglycemia) model for loss of β cell identity in diabetes.</p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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'description' => '<p>The early endosperm tissue of dicot species is very difficult to isolate by manual dissection. This protocol details how to apply the INTACT (isolation of nuclei tagged in specific cell types) system for isolating early endosperm nuclei of Arabidopsis at high purity and how to generate parental-specific epigenome profiles. As a Protocol Extension, this article describes an adaptation of an existing Nature Protocol that details the use of the INTACT method for purification of root nuclei. We address how to obtain the INTACT lines, generate the starting material and purify the nuclei. We describe a method that allows purity assessment, which has not been previously addressed. The purified nuclei can be used for ChIP and DNA bisulfite treatment followed by next-generation sequencing (seq) to study histone modifications and DNA methylation profiles, respectively. By using two different Arabidopsis accessions as parents that differ by a large number of single-nucleotide polymorphisms (SNPs), we were able to distinguish the parental origin of epigenetic modifications. Our protocol describes the only working method to our knowledge for generating parental-specific epigenome profiles of the early Arabidopsis endosperm. The complete protocol, from silique collection to finished libraries, can be completed in 2 d for bisulfite-seq (BS-seq) and 3 to 4 d for ChIP-seq experiments.This protocol is an extension to: Nat. Protoc. 6, 56-68 (2011); doi:10.1038/nprot.2010.175; published online 16 December 2010.</p>',
'date' => '2017-02-01',
'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/28055034',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<div class="row">
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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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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<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>
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<li><strong>Classified</strong> based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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'authors' => 'Lu TT, Heyne S, Dror E, Casas E, Leonhardt L, Boenke T, Yang CH, Sagar , Arrigoni L, Dalgaard K, Teperino R, Enders L, Selvaraj M, Ruf M, Raja SJ, Xie H, Boenisch U, Orkin SH, Lynn FC, Hoffman BG, Grün D, Vavouri T, Lempradl AM, Pospisilik JA',
'description' => '<p>To date, it remains largely unclear to what extent chromatin machinery contributes to the susceptibility and progression of complex diseases. Here, we combine deep epigenome mapping with single-cell transcriptomics to mine for evidence of chromatin dysregulation in type 2 diabetes. We find two chromatin-state signatures that track β cell dysfunction in mice and humans: ectopic activation of bivalent Polycomb-silenced domains and loss of expression at an epigenomically unique class of lineage-defining genes. β cell-specific Polycomb (Eed/PRC2) loss of function in mice triggers diabetes-mimicking transcriptional signatures and highly penetrant, hyperglycemia-independent dedifferentiation, indicating that PRC2 dysregulation contributes to disease. The work provides novel resources for exploring β cell transcriptional regulation and identifies PRC2 as necessary for long-term maintenance of β cell identity. Importantly, the data suggest a two-hit (chromatin and hyperglycemia) model for loss of β cell identity in diabetes.</p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-DotBlot.jpg" alt="H3K27me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ELISA.jpg" alt="H3K27me1 Antibody validated in ELISA" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-DotBlot.jpg" alt="H3K27me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<div class="row">
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<div class="extra-spaced"></div>
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<div class="extra-spaced"></div>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>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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<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>To date, it remains largely unclear to what extent chromatin machinery contributes to the susceptibility and progression of complex diseases. Here, we combine deep epigenome mapping with single-cell transcriptomics to mine for evidence of chromatin dysregulation in type 2 diabetes. We find two chromatin-state signatures that track β cell dysfunction in mice and humans: ectopic activation of bivalent Polycomb-silenced domains and loss of expression at an epigenomically unique class of lineage-defining genes. β cell-specific Polycomb (Eed/PRC2) loss of function in mice triggers diabetes-mimicking transcriptional signatures and highly penetrant, hyperglycemia-independent dedifferentiation, indicating that PRC2 dysregulation contributes to disease. The work provides novel resources for exploring β cell transcriptional regulation and identifies PRC2 as necessary for long-term maintenance of β cell identity. Importantly, the data suggest a two-hit (chromatin and hyperglycemia) model for loss of β cell identity in diabetes.</p>',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<td>ChIP <sup>*</sup></td>
<td>1 µg/ChIP</td>
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<td>1:500</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.</small></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-DotBlot.jpg" alt="H3K27me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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'reactivity' => 'Human, Arabidopsis: positive. Other species: not tested.',
'type' => 'Polyclonal',
'purity' => 'Affinity purified polyclonal antibody.',
'classification' => 'Classic',
'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP <sup>*</sup></td>
<td>1 µg/ChIP</td>
<td>Fig 1</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:500</td>
<td>Fig 2</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:20,000</td>
<td>Fig 3</td>
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<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 4</td>
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<tr>
<td>Immunofluorescence</td>
<td>1:1,000</td>
<td>Fig 5</td>
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<p></p>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.</small></p>',
'storage_conditions' => 'Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.',
'storage_buffer' => 'PBS containing 0.05% azide and 0.05% ProClin 300.',
'precautions' => 'This product is for research use only. Not for use in diagnostic or therapeutic procedures.',
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'name' => 'H3K27me1 Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the monomethylated lysine 27 (<strong>H3K27me1</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation data',
'info1' => '<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP-b.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ELISA.jpg" alt="H3K27me1 Antibody validated in ELISA" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-DotBlot.jpg" alt="H3K27me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
</div>
</div>
<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
</div>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
<div class="small-12 medium-6 large-6 columns">
<p></p>
<p></p>
<p></p>
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<p></p>
<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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'meta_description' => 'Diagenode Offers Extensively Validated ChIP-Grade Antibodies, Confirmed for their Specificity, and high level of Performance in Chromatin Immunoprecipitation ChIP',
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'description' => '<h1><strong>Validated epigenetics antibodies</strong> – care for a sample?<br /> </h1>
<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
<ul>
<li><strong>Focused</strong> - Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
<li><strong>Strict quality standards</strong> with rigorous QC and validation</li>
<li><strong>Classified</strong> based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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'description' => '<p>To date, it remains largely unclear to what extent chromatin machinery contributes to the susceptibility and progression of complex diseases. Here, we combine deep epigenome mapping with single-cell transcriptomics to mine for evidence of chromatin dysregulation in type 2 diabetes. We find two chromatin-state signatures that track β cell dysfunction in mice and humans: ectopic activation of bivalent Polycomb-silenced domains and loss of expression at an epigenomically unique class of lineage-defining genes. β cell-specific Polycomb (Eed/PRC2) loss of function in mice triggers diabetes-mimicking transcriptional signatures and highly penetrant, hyperglycemia-independent dedifferentiation, indicating that PRC2 dysregulation contributes to disease. The work provides novel resources for exploring β cell transcriptional regulation and identifies PRC2 as necessary for long-term maintenance of β cell identity. Importantly, the data suggest a two-hit (chromatin and hyperglycemia) model for loss of β cell identity in diabetes.</p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-ChIP.png" alt="H3K27me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K27me1</strong><br />ChIP was performed with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) on sheared chromatin from 500,000 K562 cells using the “iDeal ChIP-seq” kit (Cat. No. C01010051). The chromatin was spiked with a panel of in vitro assembled nucleosomes, each containing a specific lysine methylation (SNAP-ChIP K-MetStat Panel, Epicypher). A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control.<br /> <strong>Figure 1A.</strong> Quantitative PCR was performed with primers for the coding sequence of the active GAPDH and ACTB genes, used as positive controls, and for the inactive MYOD1 and MYT1 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).<br /> <strong>Figure 1B.</strong> Recovery of the nucleosomes carrying the H3K27me1, H3K27me2, H3K27me3, H3K4me1, H3K9me1, H3K36me1, H4K20me1 and the unmodified H3K4 as determined by qPCR. The figure clearly shows the antibody is specific in ChIP for the H3K27me1 modification with some slight cross reaction with H3K36me1 and H3K9me1 at higher concentrations. </small></p>
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<p><small><strong>Figure 2. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3K27me1 (Cat. No. C15410045). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1:32,900.</small></p>
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<p><small><strong>Figure 3. Cross reactivity tests using the Diagenode antibody directed against H3K27me1</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K27me1 (Cat. No. C15410045) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest.</small></p>
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<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410045-WB.png" alt="H3K27me1 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K27me1</strong><br />Histone extracts (15 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against H3K27me1 (Cat. No. C15410045) 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.</small></p>
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<div class="row">
<div class="small-6 columns">
<p>A. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-A.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>B. <img src="https://www.diagenode.com/img/product/antibodies/C15410045_IF-B.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>C. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-C.jpg" alt="H3K27me1 Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<p>D. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-D.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p>E. <img src="https://www.diagenode.com/img/product/antibodies/C15410045-IF-E.jpg" alt="H3K27me1 Antibody validated for Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K27me1</strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3K27me1 (Cat. No. C15410045) and with DAPI. Cells were fixed with 4% formaldehyde for 20’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 5A: cells were immunofluorescently labeled with the H3K27me1 antibody (left) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C, D and E: staining of the cells with the H3K27me1 antibody after incubation of the antibody with 2 ng/µl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K27, respectively.</small></p>
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'description' => '<p>The early endosperm tissue of dicot species is very difficult to isolate by manual dissection. This protocol details how to apply the INTACT (isolation of nuclei tagged in specific cell types) system for isolating early endosperm nuclei of Arabidopsis at high purity and how to generate parental-specific epigenome profiles. As a Protocol Extension, this article describes an adaptation of an existing Nature Protocol that details the use of the INTACT method for purification of root nuclei. We address how to obtain the INTACT lines, generate the starting material and purify the nuclei. We describe a method that allows purity assessment, which has not been previously addressed. The purified nuclei can be used for ChIP and DNA bisulfite treatment followed by next-generation sequencing (seq) to study histone modifications and DNA methylation profiles, respectively. By using two different Arabidopsis accessions as parents that differ by a large number of single-nucleotide polymorphisms (SNPs), we were able to distinguish the parental origin of epigenetic modifications. Our protocol describes the only working method to our knowledge for generating parental-specific epigenome profiles of the early Arabidopsis endosperm. The complete protocol, from silique collection to finished libraries, can be completed in 2 d for bisulfite-seq (BS-seq) and 3 to 4 d for ChIP-seq experiments.This protocol is an extension to: Nat. Protoc. 6, 56-68 (2011); doi:10.1038/nprot.2010.175; published online 16 December 2010.</p>',
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