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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-ChIP.png" alt="H3T6pK9me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. H3T6pK9me1 antibody ChIP results</strong><br /> Chromatin Immunoprecipitation of H3T6pK9me1 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of H3T6pK9me1 and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-IF.png" alt="H3T6pK9me1 Antibody validated in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2. H3T6pK9me1 antibody Immunofluorescence results</strong><br /> Immunofluorescence of H3T6pK9me1 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: incubated at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10,000 for 45 min at RT. Localization: H3T6pK9me1 is nuclear and chromosomal. Staining: H3T6pK9me1 is expressed in green and the nuclei are counterstained with DAPI (blue). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB.jpg" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="74" height="185" /></p>
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<p><small><strong> Figure 3. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg NIH-3T3 Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-2.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="72" height="180" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg C. elegans embryo lysate. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-3.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="81" height="191" /></p>
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<p><small><strong> Figure 5. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg HeLa Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-DotBlot.png" alt="H3T6pK9me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 6. H3T6pK9me1 antibody Dot blot results</strong><br /> Dot Blot of H3T6pK9me1 antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody diluted 1:1,000 for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT. </small></p>
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<td>1:200</td>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-ChIP.png" alt="H3T6pK9me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. H3T6pK9me1 antibody ChIP results</strong><br /> Chromatin Immunoprecipitation of H3T6pK9me1 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of H3T6pK9me1 and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-IF.png" alt="H3T6pK9me1 Antibody validated in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2. H3T6pK9me1 antibody Immunofluorescence results</strong><br /> Immunofluorescence of H3T6pK9me1 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: incubated at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10,000 for 45 min at RT. Localization: H3T6pK9me1 is nuclear and chromosomal. Staining: H3T6pK9me1 is expressed in green and the nuclei are counterstained with DAPI (blue). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB.jpg" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="74" height="185" /></p>
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<p><small><strong> Figure 3. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg NIH-3T3 Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 4. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg C. elegans embryo lysate. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 5. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg HeLa Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-DotBlot.png" alt="H3T6pK9me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 6. H3T6pK9me1 antibody Dot blot results</strong><br /> Dot Blot of H3T6pK9me1 antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody diluted 1:1,000 for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><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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'meta_description' => 'H3T6pK9me1 (Histone H3 monomethylated at lysine 9 and phosphorylated at threonin 6) Polyclonal Antibody validated in ChIP-qPCR, IHC, WB, IF and DB. ',
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'description' => '<p><span>Polyclonal antibody raised in rabbit against Histone H3 (p Thr6, monomethyl Lys9), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-ChIP.png" alt="H3T6pK9me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1. H3T6pK9me1 antibody ChIP results</strong><br /> Chromatin Immunoprecipitation of H3T6pK9me1 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of H3T6pK9me1 and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-IF.png" alt="H3T6pK9me1 Antibody validated in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 2. H3T6pK9me1 antibody Immunofluorescence results</strong><br /> Immunofluorescence of H3T6pK9me1 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: incubated at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10,000 for 45 min at RT. Localization: H3T6pK9me1 is nuclear and chromosomal. Staining: H3T6pK9me1 is expressed in green and the nuclei are counterstained with DAPI (blue). </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB.jpg" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="74" height="185" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg NIH-3T3 Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-2.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="72" height="180" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg C. elegans embryo lysate. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-3.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="81" height="191" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg HeLa Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-DotBlot.png" alt="H3T6pK9me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 6. H3T6pK9me1 antibody Dot blot results</strong><br /> Dot Blot of H3T6pK9me1 antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody diluted 1:1,000 for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT. </small></p>
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'description' => 'Methylation of Histone H3 at Lys9 (K9) is an epigenetic silencer of transcription. Gene silencing from histone post translational modifications, as well as DNA methylation, play a key role in the development of normal tissues. If this silencing is disturbed through the artificial silencing of RIZ1, and thereby H3 K9Me1, it has been shown that normal apoptotic processes in precancerous cells can be reduced. Interestingly, data indicates that the conversion of the monomethyl to the trimethyl form requires mediation by SUVR4 in transposons and pseudogenes. Research also indicates that the presence of the G9a/GLP heterodimeric complex is required for this modification to exist. The additional phosphorylation at Thr6 (T6p) affects the ability of other proteins to bind to the H3 tail, along with amplifing the effects of other histone PTMs that are present. Because T6 phosphorylation is constitutive, its dephosphorylation may play a key role in DNA transcription, repair and replication.',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-ChIP.png" alt="H3T6pK9me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. H3T6pK9me1 antibody ChIP results</strong><br /> Chromatin Immunoprecipitation of H3T6pK9me1 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of H3T6pK9me1 and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 2. H3T6pK9me1 antibody Immunofluorescence results</strong><br /> Immunofluorescence of H3T6pK9me1 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: incubated at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10,000 for 45 min at RT. Localization: H3T6pK9me1 is nuclear and chromosomal. Staining: H3T6pK9me1 is expressed in green and the nuclei are counterstained with DAPI (blue). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB.jpg" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="74" height="185" /></p>
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<p><small><strong> Figure 3. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg NIH-3T3 Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-2.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="72" height="180" /></p>
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<p><small><strong> Figure 4. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg C. elegans embryo lysate. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-3.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="81" height="191" /></p>
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<p><small><strong> Figure 5. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg HeLa Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-DotBlot.png" alt="H3T6pK9me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 6. H3T6pK9me1 antibody Dot blot results</strong><br /> Dot Blot of H3T6pK9me1 antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody diluted 1:1,000 for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT. </small></p>
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<td>2-5 μg/million cells</td>
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><small><strong> Figure 1. H3T6pK9me1 antibody ChIP results</strong><br /> Chromatin Immunoprecipitation of H3T6pK9me1 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of H3T6pK9me1 and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 2. H3T6pK9me1 antibody Immunofluorescence results</strong><br /> Immunofluorescence of H3T6pK9me1 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: incubated at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10,000 for 45 min at RT. Localization: H3T6pK9me1 is nuclear and chromosomal. Staining: H3T6pK9me1 is expressed in green and the nuclei are counterstained with DAPI (blue). </small></p>
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<p><small><strong> Figure 3. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg NIH-3T3 Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 4. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg C. elegans embryo lysate. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-3.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="81" height="191" /></p>
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<p><small><strong> Figure 5. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg HeLa Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 6. H3T6pK9me1 antibody Dot blot results</strong><br /> Dot Blot of H3T6pK9me1 antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody diluted 1:1,000 for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT. </small></p>
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<p><small><strong> Figure 2. H3T6pK9me1 antibody Immunofluorescence results</strong><br /> Immunofluorescence of H3T6pK9me1 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: incubated at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10,000 for 45 min at RT. Localization: H3T6pK9me1 is nuclear and chromosomal. Staining: H3T6pK9me1 is expressed in green and the nuclei are counterstained with DAPI (blue). </small></p>
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<p><small><strong> Figure 3. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg NIH-3T3 Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 4. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg C. elegans embryo lysate. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 5. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg HeLa Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 6. H3T6pK9me1 antibody Dot blot results</strong><br /> Dot Blot of H3T6pK9me1 antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody diluted 1:1,000 for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT. </small></p>
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'description' => 'Methylation of Histone H3 at Lys9 (K9) is an epigenetic silencer of transcription. Gene silencing from histone post translational modifications, as well as DNA methylation, play a key role in the development of normal tissues. If this silencing is disturbed through the artificial silencing of RIZ1, and thereby H3 K9Me1, it has been shown that normal apoptotic processes in precancerous cells can be reduced. Interestingly, data indicates that the conversion of the monomethyl to the trimethyl form requires mediation by SUVR4 in transposons and pseudogenes. Research also indicates that the presence of the G9a/GLP heterodimeric complex is required for this modification to exist. The additional phosphorylation at Thr6 (T6p) affects the ability of other proteins to bind to the H3 tail, along with amplifing the effects of other histone PTMs that are present. Because T6 phosphorylation is constitutive, its dephosphorylation may play a key role in DNA transcription, repair and replication.',
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<th>Suggested dilution</th>
<th>References</th>
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<td>ChIP</td>
<td>2-5 μg/million cells</td>
<td>Fig 1</td>
</tr>
<tr>
<td>Immunohistochemistry</td>
<td>1:200</td>
<td></td>
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<td>Immunofluorescence</td>
<td>1:100</td>
<td>Fig 2</td>
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<td>Western Blotting</td>
<td>1:500 - 1:1,000</td>
<td>Fig 3, 4, 5</td>
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<td>Dot Blotting</td>
<td>1:1,000</td>
<td>Fig 6</td>
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><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></p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><small><strong> Figure 1. H3T6pK9me1 antibody ChIP results</strong><br /> Chromatin Immunoprecipitation of H3T6pK9me1 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of H3T6pK9me1 and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 2. H3T6pK9me1 antibody Immunofluorescence results</strong><br /> Immunofluorescence of H3T6pK9me1 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: incubated at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10,000 for 45 min at RT. Localization: H3T6pK9me1 is nuclear and chromosomal. Staining: H3T6pK9me1 is expressed in green and the nuclei are counterstained with DAPI (blue). </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB.jpg" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="74" height="185" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg NIH-3T3 Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-2.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="72" height="180" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg C. elegans embryo lysate. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-WB-3.png" alt="H3T6pK9me1 Antibody validated in Western Blot " style="display: block; margin-left: auto; margin-right: auto;" width="81" height="191" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg HeLa Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-DotBlot.png" alt="H3T6pK9me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 6. H3T6pK9me1 antibody Dot blot results</strong><br /> Dot Blot of H3T6pK9me1 antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody diluted 1:1,000 for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-ChIP.png" alt="H3T6pK9me1 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. H3T6pK9me1 antibody ChIP results</strong><br /> Chromatin Immunoprecipitation of H3T6pK9me1 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2ug of H3T6pK9me1 and 20ul of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using quantitative real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 2. H3T6pK9me1 antibody Immunofluorescence results</strong><br /> Immunofluorescence of H3T6pK9me1 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody: incubated at a 1:100 dilution for 1 h at RT. Secondary antibody: FITC secondary antibody at 1:10,000 for 45 min at RT. Localization: H3T6pK9me1 is nuclear and chromosomal. Staining: H3T6pK9me1 is expressed in green and the nuclei are counterstained with DAPI (blue). </small></p>
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<p><small><strong> Figure 3. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg NIH-3T3 Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 4. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg C. elegans embryo lysate. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><small><strong> Figure 5. H3T6pK9me1 antibody Western blot results</strong><br /> Western Blot of H3T6pK9me1 antibody. 30 μg HeLa Histone extracts. Primary antibody diluted 1:1,000 overnight at 4°C. Secondary antibody: IRDye800TM rabbit secondary antibody at 1:10,000 for 45 min at RT. Predicted/Observed size: ~15 kDa. Other band(s): None. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410283-DotBlot.png" alt="H3T6pK9me1 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 6. H3T6pK9me1 antibody Dot blot results</strong><br /> Dot Blot of H3T6pK9me1 antibody. Lane 1: Histone H3 1-13. Lane 2: T6p. Lane 3: T6pK9Me1. Lane 4: T6pK9Me2. Lane 5: T6pK9Me3. Lane 6: Histone H3 2-17. Lane 7: K9Me1. Lane 8: K9Me2. Lane 9: K9Me3. Lane 10: K9Ac. Load: 1, 10, and 100 picomoles of peptide. Primary antibody diluted 1:1,000 for 45 min at 4°C. Secondary antibody: DylightTM488 rabbit secondary antibody at 1:10,000 for 45 min at RT. </small></p>
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<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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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>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>
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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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