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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1-bis.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<td>ChIP <sup>*</sup></td>
<td>2 μg/ChIP</td>
<td>Fig 1</td>
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<td>1:100</td>
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<td>1:20,000</td>
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<td>1:1,000</td>
<td>Fig 4</td>
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<td>1:2,000</td>
<td>Fig 5</td>
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'name' => 'H3S10p Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the phosphorylated serine 10 (<strong>H3S10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1-bis.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<td>2 μg/ChIP</td>
<td>Fig 1</td>
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<td>1:100</td>
<td>Fig 2</td>
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<td>1:20,000</td>
<td>Fig 3</td>
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<td>1:1,000</td>
<td>Fig 4</td>
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<td>Fig 5</td>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1-bis.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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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><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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'description' => '<p>We investigated the relationship among ERK signaling, histone modifications, and transcription factor activity, focusing on the ERK-regulated ternary complex factor family of SRF partner proteins. In MEFs, activation of ERK by TPA stimulation induced a common pattern of H3K9acS10ph, H4K16ac, H3K27ac, H3K9acK14ac, and H3K4me3 at hundreds of transcription start site (TSS) regions and remote regulatory sites. The magnitude of the increase in histone modification correlated well with changes in transcription. H3K9acS10ph preceded the other modifications. Most induced changes were TCF dependent, but TCF-independent TSSs exhibited the same hierarchy, indicating that it reflects gene activation per se. Studies with TCF Elk-1 mutants showed that TCF-dependent ERK-induced histone modifications required Elk-1 to be phosphorylated and competent to activate transcription. Analysis of direct TCF-SRF target genes and chromatin modifiers confirmed this and showed that H3S10ph required only Elk-1 phosphorylation. Induction of histone modifications following ERK stimulation is thus directed by transcription factor activation and transcription.</p>',
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<thead>
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<th>References</th>
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<td>ChIP <sup>*</sup></td>
<td>2 μg/ChIP</td>
<td>Fig 1</td>
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<tr>
<td>ELISA</td>
<td>1:100</td>
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<td>Dot Blotting</td>
<td>1:20,000</td>
<td>Fig 3</td>
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<td>1:1,000</td>
<td>Fig 4</td>
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<td>Fig 5</td>
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<p><small><sup>*</sup> Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
</div>
</div>',
'label2' => 'Target Description',
'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. Phosphorylation of H3S10 is associated with mitosis.</p>',
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'catalog_number' => 'C15410116',
'old_catalog_number' => 'pAb-116-050',
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'type' => 'FRE',
'search_order' => '03-Antibody',
'price_EUR' => '380',
'price_USD' => '380',
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'price_JPY' => '59525',
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'slug' => 'h3s10p-polyclonal-antibody-classic-50-mg-48-ml',
'meta_title' => 'H3S10p Antibody - ChIP Grade (C15410116) | Diagenode',
'meta_keywords' => '',
'meta_description' => 'H3S10p (Histone H3 phosphorylated at serine 10) Polyclonal Antibody validated in ChIP-qPCR, ELISA, DB, WB and IF. Batch-specific data available on the website. Sample size available.',
'modified' => '2024-11-19 17:01:16',
'created' => '2015-06-29 14:08:20',
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'name' => 'H3S10p polyclonal antibody',
'description' => 'Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Phosphorylation of H3S10 is associated with mitosis.',
'clonality' => '',
'isotype' => '',
'lot' => 'A160-001234P',
'concentration' => '1.05 µg/µl',
'reactivity' => 'Human',
'type' => 'Polyclonal',
'purity' => 'Affinity purified',
'classification' => 'Classic',
'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP <sup>*</sup></td>
<td>2 μg/ChIP</td>
<td>Fig 1</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:100</td>
<td>Fig 2</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:20,000</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 4</td>
</tr>
<tr>
<td>Immunofluorescence</td>
<td>1:2,000</td>
<td>Fig 5</td>
</tr>
</tbody>
</table>
<p><small><sup>*</sup> Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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'name' => 'H3S10p Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the phosphorylated serine 10 (<strong>H3S10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1-bis.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
</div>
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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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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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'meta_description' => 'Polyclonal and Monoclonal Antibodies against Histones and their modifications validated for many applications, including Chromatin Immunoprecipitation (ChIP) and ChIP-Sequencing (ChIP-seq)',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</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><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="Immunofluorescence figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="Immunofluorescence figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="Western blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="Immunofluorescence figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="Immunofluorescence figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="Immunofluorescence figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="Immunofluorescence figure D" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the phosphorylated serine 10 (H3S10p), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="ChIP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1-bis.jpg" alt="ChIP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="ELISA" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="Western blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="Immunofluorescence figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="Immunofluorescence figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="Immunofluorescence figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="Immunofluorescence figure D" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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'meta_description' => 'H3S10p (Histone H3 phosphorylated at serine 10) Polyclonal Antibody validated in ChIP-qPCR, ELISA, DB, WB and IF. Batch-specific data available on the website. Sample size available.',
'meta_title' => 'H3S10p Antibody - ChIP Grade (C15410116) | Diagenode',
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'id' => '2248',
'antibody_id' => '179',
'name' => 'H3S10p Antibody',
'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the phosphorylated serine 10 (<strong>H3S10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1-bis.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
</div>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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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. Phosphorylation of H3S10 is associated with mitosis.</p>',
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'name' => 'H3S10p polyclonal antibody',
'description' => 'Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called “histone code”. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases. Phosphorylation of H3S10 is associated with mitosis.',
'clonality' => '',
'isotype' => '',
'lot' => 'A160-001234P',
'concentration' => '1.05 µg/µl',
'reactivity' => 'Human',
'type' => 'Polyclonal',
'purity' => 'Affinity purified',
'classification' => 'Classic',
'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP <sup>*</sup></td>
<td>2 μg/ChIP</td>
<td>Fig 1</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:100</td>
<td>Fig 2</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:20,000</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 4</td>
</tr>
<tr>
<td>Immunofluorescence</td>
<td>1:2,000</td>
<td>Fig 5</td>
</tr>
</tbody>
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<p><small><sup>*</sup> Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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'application_table' => '<table>
<thead>
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<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
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<td>ChIP <sup>*</sup></td>
<td>2 μg/ChIP</td>
<td>Fig 1</td>
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<tr>
<td>ELISA</td>
<td>1:100</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>1:1,000</td>
<td>Fig 4</td>
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<tr>
<td>Immunofluorescence</td>
<td>1:2,000</td>
<td>Fig 5</td>
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<p><small><sup>*</sup> Please note that of the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the phosphorylated serine 10 (H3S10p), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="ChIP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1-bis.jpg" alt="ChIP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="ELISA" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="Western blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="Immunofluorescence figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="Immunofluorescence figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="Immunofluorescence figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="Immunofluorescence figure D" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="ChIP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="ELISA" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="Western blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="Immunofluorescence figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="Immunofluorescence figure B" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="Immunofluorescence figure C" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="Immunofluorescence figure D" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="Western blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="Immunofluorescence figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="Immunofluorescence figure D" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<td>2 μg/ChIP</td>
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'description' => '<p><span>Polyclonal antibody raised in rabbit against histone H3 containing the phosphorylated serine 10 (<strong>H3S10p</strong>), using a KLH-conjugated synthetic peptide.</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1-bis.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<td>1:1,000</td>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig1.jpg" alt="H3S10p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig2.jpg" alt="H3S10p Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig3.jpg" alt="H3S10p Antibody valiadted in Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
</div>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116_fig4.jpg" alt="H3S10p Antibody valiadted in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-B.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-C.jpg" alt="H3S10p Antibody valiadted in Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-D.jpg" alt="H3S10p Antibody valiadted for Immunofluorescence " style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>”, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><small><strong> Figure 1 ChIP results obtained with the Diagenode antibody directed against H3S10p</strong><br /> Figure 1A: ChIP assays were performed using human HeLa cells treated with colcemid to block the cells in metaphase, the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and optimized PCR primer sets for qPCR. ChIP was performed with the “LowCell# ChIP” kit (Cat. No. kch-maglow-016) on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 μg per ChIP experiment was analysed. IgG (5 μg/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos (Cat. No. pp-1004-050) and RPL30. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure 1B: ChIP was performed as described above using 2 μg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><small><strong> Figure 3 Cross reactivity test with the Diagenode antibody directed against H3S10p</strong><br /> A Dot Blot analysis was performed to test the cross reactivity of the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) with peptides containing other modifications of histone H3 or the unmodified H3S10 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated. </small></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410116-IF-A.jpg" alt="Immunofluorescence figure A" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 5 Immunofluorescence using the Diagenode antibody directed against H3S10p</strong><br /> Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) and with DAPI. Cells were fixed with 3.7% formaldehyde in PBS for 20’ at RT, followed by a 20’ permeabilization with 0.5% Triton X-100 in PBS and blocked PBS/TX-100 containing 5% normal goat serum Figure 5A: cells were immunofluorescently labeled with the H3S10p antibody (left), diluted 1:2,000 in blocking buffer followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure 5B, C and D: staining of the cells with the H3S10p antibody after incubation of the antibody with 2 μM blocking peptide containing the unmodified H3S10 sequence, the phosphorylated H3S10 and the phosphorylated H3T11, respectively. </small></p>
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<p><small><strong> Figure 2 Determination of the antibody titer</strong><br /> To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H3S10p (Cat. No. pAb-116-050) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:5,200. </small></p>
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<p><small><strong> Figure 4 Western blot analysis using the Diagenode antibody directed against H3S10p</strong><br /> HeLa cells were treated with colcemid, which blocks the cell cycle in metaphase and 15 μg of histone extracts of the cells were analysed by Western blot using the Diagenode antibody against H3S10p (Cat. No. pAb-116-050) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. </small></p>
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'slug' => 'datasheet-h3s10p-pab-116-050',
'meta_keywords' => null,
'meta_description' => null,
'modified' => '2015-07-07 11:47:44',
'created' => '2015-07-07 11:47:44',
'ProductsDocument' => array(
'id' => '488',
'product_id' => '2248',
'document_id' => '715'
)
)
$sds = array(
'id' => '3518',
'name' => 'SDS C15410116 H3S10p Antibody DE de',
'language' => 'de',
'url' => 'files/SDS/H3S10p/SDS-C15410116-H3S10p_Antibody-DE-de-GHS_2_0.pdf',
'countries' => 'DE',
'modified' => '2024-01-16 16:02:59',
'created' => '2024-01-16 16:02:59',
'ProductsSafetySheet' => array(
'id' => '5728',
'product_id' => '2248',
'safety_sheet_id' => '3518'
)
)
$publication = array(
'id' => '3174',
'name' => 'ERK-Induced Activation of TCF Family of SRF Cofactors Initiates a Chromatin Modification Cascade Associated with Transcription',
'authors' => 'Esnault C. et al.',
'description' => '<p>We investigated the relationship among ERK signaling, histone modifications, and transcription factor activity, focusing on the ERK-regulated ternary complex factor family of SRF partner proteins. In MEFs, activation of ERK by TPA stimulation induced a common pattern of H3K9acS10ph, H4K16ac, H3K27ac, H3K9acK14ac, and H3K4me3 at hundreds of transcription start site (TSS) regions and remote regulatory sites. The magnitude of the increase in histone modification correlated well with changes in transcription. H3K9acS10ph preceded the other modifications. Most induced changes were TCF dependent, but TCF-independent TSSs exhibited the same hierarchy, indicating that it reflects gene activation per se. Studies with TCF Elk-1 mutants showed that TCF-dependent ERK-induced histone modifications required Elk-1 to be phosphorylated and competent to activate transcription. Analysis of direct TCF-SRF target genes and chromatin modifiers confirmed this and showed that H3S10ph required only Elk-1 phosphorylation. Induction of histone modifications following ERK stimulation is thus directed by transcription factor activation and transcription.</p>',
'date' => '2017-03-09',
'pmid' => 'http://www.cell.com/molecular-cell/abstract/S1097-2765(17)30097-7',
'doi' => '',
'modified' => '2017-05-10 16:44:53',
'created' => '2017-05-10 16:44:53',
'ProductsPublication' => array(
'id' => '2007',
'product_id' => '2248',
'publication_id' => '3174'
)
)
$externalLink = ' <a href="http://www.cell.com/molecular-cell/abstract/S1097-2765(17)30097-7" target="_blank"><i class="fa fa-external-link"></i></a>'include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118
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