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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p>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>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against <strong>histone H3</strong> containing the <strong>acetylated lysine 9 and the phosphorylated serine 10</strong> (<strong>H3K9acS10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_chip.png" alt="H3K9acS10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="396" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_ELISA.png" alt="H3K9acS10p Antibody ELISA validation" caption="false" width="400" height="316" /></p>
<p class="text-center"></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_DotBlot.png" alt="H3K9acS10p Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" width="700" height="310" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_WB.png" alt="H3K9acS10p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" width="186" height="223" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_DotBlot.png" alt="H3K9acS10p Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" width="700" height="310" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_WB.png" alt="H3K9acS10p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" width="186" height="223" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_IF.png" alt="H3K9acS10p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="99" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>',
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'info2' => '<p>Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Acetylation of K9 and phosphorylation of S10 of histone H3 are associated with active gene transcription</p>',
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'lot' => 'A397-001 ',
'concentration' => 'not determined',
'reactivity' => 'Human',
'type' => 'Polyclonal',
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'application_table' => '<table>
<thead>
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<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
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<td>ChIP <sup>*</sup></td>
<td>15 μl/ChIP</td>
<td>Fig 1</td>
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<tr>
<td>ELISA</td>
<td>1:1,000 - 1:4,000</td>
<td>Fig 2</td>
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<tr>
<td>Dot Blotting</td>
<td>1:20,000</td>
<td>Fig 3</td>
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<tr>
<td>Western Blotting</td>
<td>1:250</td>
<td>Fig 4</td>
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<tr>
<td>IF</td>
<td>1:500</td>
<td>Fig 5</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-10 μl per IP.</small></p>',
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'name' => 'H3K9acS10p Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against <strong>histone H3</strong> containing the <strong>acetylated lysine 9 and the phosphorylated serine 10</strong> (<strong>H3K9acS10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_chip.png" alt="H3K9acS10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="396" /></p>
<p></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_ELISA.png" alt="H3K9acS10p Antibody ELISA validation" caption="false" width="400" height="316" /></p>
<p class="text-center"></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_DotBlot.png" alt="H3K9acS10p Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" width="700" height="310" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_WB.png" alt="H3K9acS10p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" width="186" height="223" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_IF.png" alt="H3K9acS10p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="99" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against <strong>histone H3</strong> containing the <strong>acetylated lysine 9 and the phosphorylated serine 10</strong> (<strong>H3K9acS10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_chip.png" alt="H3K9acS10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="396" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_ELISA.png" alt="H3K9acS10p Antibody ELISA validation" caption="false" width="400" height="316" /></p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_ELISA.png" alt="H3K9acS10p Antibody ELISA validation" caption="false" width="400" height="316" /></p>
<p class="text-center"></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_DotBlot.png" alt="H3K9acS10p Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" width="700" height="310" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
</div>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_WB.png" alt="H3K9acS10p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" width="186" height="223" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_IF.png" alt="H3K9acS10p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="99" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p>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>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against <strong>histone H3</strong> containing the <strong>acetylated lysine 9 and the phosphorylated serine 10</strong> (<strong>H3K9acS10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_chip.png" alt="H3K9acS10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="396" /></p>
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<div class="small-6 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_ELISA.png" alt="H3K9acS10p Antibody ELISA validation" caption="false" width="400" height="316" /></p>
<p class="text-center"></p>
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<div class="small-6 columns">
<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_DotBlot.png" alt="H3K9acS10p Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" width="700" height="310" /></p>
</div>
<div class="small-6 columns">
<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<div class="row">
<div class="small-6 columns">
<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_WB.png" alt="H3K9acS10p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" width="186" height="223" /></p>
</div>
<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_IF.png" alt="H3K9acS10p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="99" /></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against <strong>histone H3</strong> containing the <strong>acetylated lysine 9 and the phosphorylated serine 10</strong> (<strong>H3K9acS10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_chip.png" alt="H3K9acS10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="396" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against H3K9acS10p</strong><br />ChIP assays were performed using human osteosarcoma (U2OS) cells, the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) and optimized PCR primer sets for qPCR. Chromatin was sheared with the Diagenode “Shearing ChIP” kit (Cat. No. kch-redmod-100). ChIP was performed with the “OneDay ChIP” kit (Cat. No. kch-oneDIP-060), using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 μl per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_ELISA.png" alt="H3K9acS10p Antibody ELISA validation" caption="false" width="400" height="316" /></p>
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<p><small><strong>Figure 2. Determination of the titer</strong><br />To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody against human H3K9acS10p (Cat. No. CS-102-100). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:89,000.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_DotBlot.png" alt="H3K9acS10p Antibody validated in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" width="700" height="310" /></p>
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<p><small><strong>Figure 3. Cross reactivity test using the Diagenode antibody directed against H3K9acS10p</strong><br />A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the double modification.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_WB.png" alt="H3K9acS10p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" width="186" height="223" /></p>
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<div class="small-6 columns">
<p><small> <strong>Figure 4. Western blot analysis using the Diagenode antibody directed against H3K9acS10p</strong><br />Histone extracts of HeLa cells (15 μg) were analysed by Western blot using the Diagenode antibody against H3K9acS10p (Cat. No. CS-102-100) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.</small></p>
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<p class="text-center"><img src="https://www.diagenode.com/img/product/antibodies/C15310102_IF.png" alt="H3K9acS10p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" width="400" height="99" /></p>
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<p><small> <strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against H3K9acS10p</strong><br />HeLa cells were stained with the Diagenode antibody against H3K9acS10p (cat. No. C15310102) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9acS10p antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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