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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410291-ChIP.png" alt="H3K18me2 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. ChIP</strong><br /> Chromatin Immunoprecipitation of H3K18me2 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 2. Immunofluorescence</strong><br /> Immunofluorescence of H3K18me2 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:500 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone H3K18me2 is nuclear and chromosomal. Staining: Histone H3K18me2 is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 594 (red). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410291-DB.png" alt="H3K18me2 Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 3. Dot Blot</strong><br /> Dot Blot of H3K18me2 antibody. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1,000 dilution 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/C15410291-WB1.png" alt="H3K18me2 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</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of C. elegans embryo lysate. Primary antibody used at a 1:500 dilution 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/C15410291-WB2.png" alt="H3K18me2 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution 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/C15410291-ChIP.png" alt="H3K18me2 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. ChIP</strong><br /> Chromatin Immunoprecipitation of H3K18me2 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 2. Immunofluorescence</strong><br /> Immunofluorescence of H3K18me2 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:500 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone H3K18me2 is nuclear and chromosomal. Staining: Histone H3K18me2 is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 594 (red). </small></p>
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<p><small><strong> Figure 3. Dot Blot</strong><br /> Dot Blot of H3K18me2 antibody. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1,000 dilution 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 4. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of C. elegans embryo lysate. Primary antibody used at a 1:500 dilution 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/C15410291-WB2.png" alt="H3K18me2 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution 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>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>
</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>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p><small><strong> Figure 1. ChIP</strong><br /> Chromatin Immunoprecipitation of H3K18me2 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 3. Dot Blot</strong><br /> Dot Blot of H3K18me2 antibody. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1,000 dilution 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 4. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of C. elegans embryo lysate. Primary antibody used at a 1:500 dilution 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. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution 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 1. ChIP</strong><br /> Chromatin Immunoprecipitation of H3K18me2 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 2. Immunofluorescence</strong><br /> Immunofluorescence of H3K18me2 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:500 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone H3K18me2 is nuclear and chromosomal. Staining: Histone H3K18me2 is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 594 (red). </small></p>
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<p><small><strong> Figure 3. Dot Blot</strong><br /> Dot Blot of H3K18me2 antibody. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1,000 dilution 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 4. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of C. elegans embryo lysate. Primary antibody used at a 1:500 dilution 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. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution 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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'meta_description' => 'H3K18me2 (Histone H3 dimethylated at lysine 18) Polyclonal Antibody validated in ChIP-qPCR, IF, IHC, DB and WB. ',
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'description' => 'The di-methylated K18 on histone H3 is a seemingly transient post-translational modification. H3K18 is better known to be acetylated, and occasionally mono-methylated. Suv39h1, a well-studied histone methyltransferase seems to be responsible for the transition of acetylation and methylation at this H3 modification site. The di-methylated K18 on H3 seems to be associated with embryological development and possibly implantation.',
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<th>Suggested dilution <sup>*</sup></th>
<th>References</th>
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<tr>
<td>ChIP</td>
<td>2-5 μg/million cells</td>
<td>Fig 1</td>
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<tr>
<td>Immunofluorescence</td>
<td>1:500</td>
<td>Fig 2</td>
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<td>Western Blotting</td>
<td>1:500</td>
<td>Fig 4, 5</td>
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<td>1:100</td>
<td></td>
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'description' => '<p><strong>Immunofluorescence</strong>:</p>
<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><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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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>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' => 'H3K18me2 (Histone H3 dimethylated at lysine 18) Polyclonal Antibody validated in ChIP-qPCR, IF, IHC, DB and WB. ',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410291-ChIP.png" alt="H3K18me2 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. ChIP</strong><br /> Chromatin Immunoprecipitation of H3K18me2 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin. </small></p>
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<div class="small-8 columns">
<p><small><strong> Figure 2. Immunofluorescence</strong><br /> Immunofluorescence of H3K18me2 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:500 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone H3K18me2 is nuclear and chromosomal. Staining: Histone H3K18me2 is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 594 (red). </small></p>
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<p><small><strong> Figure 3. Dot Blot</strong><br /> Dot Blot of H3K18me2 antibody. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1,000 dilution 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/C15410291-WB1.png" alt="H3K18me2 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</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of C. elegans embryo lysate. Primary antibody used at a 1:500 dilution 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/C15410291-WB2.png" alt="H3K18me2 Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution 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/C15410291-ChIP.png" alt="H3K18me2 Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. ChIP</strong><br /> Chromatin Immunoprecipitation of H3K18me2 antibody. Chromatin from one million formaldehyde cross-linked Hela cells was used with 2 μg of H3K18me2 antibody and 20 μl of magnetic IgG beads per immunoprecipitation. A no antibody (No Ab) control was also used. Immunoprecipitated DNA was quantified using real-time PCR and normalized to the input chromatin. </small></p>
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<p><small><strong> Figure 2. Immunofluorescence</strong><br /> Immunofluorescence of H3K18me2 antibody. Tissue: HeLa cells. Fixation: 0.5% PFA. Primary antibody used at a 1:500 dilution for 1 h at RT. Secondary antibody: Dylight 488 secondary antibody at 1:10,000 for 45 min at RT. Localization: Histone H3K18me2 is nuclear and chromosomal. Staining: Histone H3K18me2 is expressed in green, nuclei and alpha-tubulin are counterstained with DAPI (blue) and Dylight 594 (red). </small></p>
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<p><small><strong> Figure 3. Dot Blot</strong><br /> Dot Blot of H3K18me2 antibody. Lane 1: K18 unmodified. Lane 2: K18Me2. Lane 3: K18Me3. Lane 4: K18ac. Lane 5: K18Me1. Load: 0.1, 0.3, and 1 picomoles of peptide. Primary antibody used at a 1:1,000 dilution 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 4. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of C. elegans embryo lysate. Primary antibody used at a 1:500 dilution 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. Western Blot</strong><br /> Western Blot of H3K18me2 antibody. 30 μg of NIH-3T3 histone extract. Primary antibody used at a 1:500 dilution 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>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>
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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>
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<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone 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><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>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>
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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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<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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