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'description' => '<p><span>Alternative names: <strong>POLR2A</strong>, <strong>RPB1</strong>, <strong>POLR2</strong>, <strong>RPOL2</strong></span></p>
<p><span>Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5 of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="spacer"></div>
<div class="spacer"></div>
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<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p></p>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<div class="spacer"></div>
<div class="row">
<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<br />
<div class="small-9 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:3,000</td>
<td>Fig 3</td>
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<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 4, 5</td>
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<td>1:500</td>
<td>Fig 6</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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'name' => 'Pol II S5p Antibody - ChIP-seq Grade',
'description' => '<p><span>Alternative names: <strong>POLR2A</strong>, <strong>RPB1</strong>, <strong>POLR2</strong>, <strong>RPOL2</strong></span></p>
<p><span>Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5 of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="spacer"></div>
<div class="spacer"></div>
<div class="row">
<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p></p>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<div class="spacer"></div>
<div class="spacer"></div>
<div class="row">
<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<br />
<div class="small-9 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<th>Suggested dilution</th>
<th>References</th>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:3,000</td>
<td>Fig 3</td>
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<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 4, 5</td>
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<td>1:500</td>
<td>Fig 6</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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'description' => '<p><span>Alternative names: <strong>POLR2A</strong>, <strong>RPB1</strong>, <strong>POLR2</strong>, <strong>RPOL2</strong></span></p>
<p><span>Monoclonal antibody raised in mouse against the <strong>YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5</strong> of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<div class="spacer"></div>
<div class="row">
<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<br />
<div class="small-9 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><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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<div class="small-10 columns">
<h3>Epigenetic antibodies you can trust!</h3>
<p>Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level of siRNA knockdown validation to assure the specificity of our non-histone antibodies.</p>
<p><strong>Short interfering RNA (siRNA)</strong> degrades target mRNA, followed by the knock-down of protein production. If the antibody that recognizes the protein of interest is specific, the Western blot of siRNA-treated cells will show a significant reduction of signal vs. untreated cells.</p>
<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
<p class="text-center"><small>WB results obtained with the HDAC1 pAb (Cat. No. C15100144) <br />on siRNA transfected cells (lane 2) and on untransfected control cells (lane 1).</small></p>
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<p><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></p>
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<p style="text-align: left;"><span style="font-weight: 400;">The below list shows our first siRNA validated antibodies. More results - coming soon</span>.</p>',
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<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>
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<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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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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<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 style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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'name' => 'CDK7 Inhibition is Effective in all the Subtypes of Breast Cancer: Determinants of Response and Synergy with EGFR Inhibition.',
'authors' => 'McDermott MSJ, Sharko AC, Munie J, Kassler S, Melendez T, Lim CU, Broude EV',
'description' => '<p>CDK7, a transcriptional cyclin-dependent kinase, is emerging as a novel cancer target. Triple-negative breast cancers (TNBC) but not estrogen receptor-positive (ER+) breast cancers have been reported to be uniquely sensitive to the CDK7 inhibitor THZ1 due to the inhibition of a cluster of TNBC-specific genes. However, bioinformatic analysis indicates that CDK7 RNA expression is associated with negative prognosis in all the major subtypes of breast cancer. To further elucidate the effects of CDK7 inhibition in breast cancer, we profiled a panel of cell lines representing different breast cancer subtypes. THZ1 inhibited cell growth in all subtypes (TNBC, HER2+, ER+, and HER2+/ER+) with no apparent subtype selectivity. THZ1 inhibited CDK7 activity and induced G1 arrest and apoptosis in all the tested cell lines, but THZ1 sensitivity did not correlate with CDK7 inhibition or CDK7 expression levels. THZ1 sensitivity across the cell line panel did not correlate with TNBC-specific gene expression but it was found to correlate with the differential inhibition of three genes: CDKN1B, MYC and transcriptional coregulator CITED2. Response to THZ1 also correlated with basal CITED2 protein expression, a potential marker of CDK7 inhibitor sensitivity. Furthermore, all of the THZ1-inhibited genes examined were inducible by EGF but THZ1 prevented this induction. THZ1 had synergistic or additive effects when combined with the EGFR inhibitor erlotinib, with no outward selectivity for a particular subtype of breast cancer. These results suggest a potential broad utility for CDK7 inhibitors in breast cancer therapy and the potential for combining CDK7 and EGFR inhibitors.</p>',
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'description' => '<p>How transcription affects genome 3D organization is not well understood. We found that during influenza A (IAV) infection, rampant transcription rapidly reorganizes host cell chromatin interactions. These changes occur at the ends of highly transcribed genes, where global inhibition of transcription termination by IAV NS1 protein causes readthrough transcription for hundreds of kilobases. In these readthrough regions, elongating RNA polymerase II disrupts chromatin interactions by inducing cohesin displacement from CTCF sites, leading to locus decompaction. Readthrough transcription into heterochromatin regions switches them from the inert (B) to the permissive (A) chromatin compartment and enables transcription factor binding. Data from non-viral transcription stimuli show that transcription similarly affects cohesin-mediated chromatin contacts within gene bodies. Conversely, inhibition of transcription elongation allows cohesin to accumulate at previously transcribed intragenic CTCF sites and to mediate chromatin looping and compaction. Our data indicate that transcription elongation by RNA polymerase II remodels genome 3D architecture.</p>',
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'description' => '<p>Although many long non-coding RNAs (lncRNAs) are imprinted, their roles often remain unknown. The Dlk1-Dio3 domain expresses the lncRNA Meg3 and multiple microRNAs and small nucleolar RNAs (snoRNAs) on the maternal chromosome and constitutes an epigenetic model for development. The domain's Dlk1 (Delta-like-1) gene encodes a ligand that inhibits Notch1 signaling and regulates diverse developmental processes. Using a hybrid embryonic stem cell (ESC) system, we find that Dlk1 becomes imprinted during neural differentiation and that this involves transcriptional upregulation on the paternal chromosome. The maternal Dlk1 gene remains poised. Its protection against activation is controlled in cis by Meg3 expression and also requires the H3-Lys-27 methyltransferase Ezh2. Maternal Meg3 expression additionally protects against de novo DNA methylation at its promoter. We find that Meg3 lncRNA is partially retained in cis and overlaps the maternal Dlk1 in embryonic cells. Combined, our data evoke an imprinting model in which allelic lncRNA expression prevents gene activation in cis.</p>',
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'description' => '<p>Of the various genetic subtypes of HIV-1, HIV-2 and SIV, only in subtype C of HIV-1, a genetically variant NF-κB binding site is found at the core of the viral promoter in association with a subtype-specific Sp1III motif. How the subtype-associated variations in the core transcription factor binding sites (TFBS) influence gene expression from the viral promoter has not been examined previously. Using panels of infectious viral molecular clones, we demonstrate that subtype-specific NF-κB and Sp1III motifs have evolved for optimal gene expression, and neither of the motifs can be substituted by a corresponding TFBS variant.The variant NF-κB motif binds NF-κB with an affinity two-fold higher than that of the generic NF-κB site. Importantly, in the context of an infectious virus, the subtype-specific Sp1III motif demonstrates a profound loss of function in association with the generic NF-κB motif. An additional substitution of the Sp1III motif fully restores viral replication suggesting that the subtype C specific Sp1III has evolved to function with the variant, but not generic, NF-κB motif. A change of only two base pairs in the central NF-κB motif completely suppresses viral transcription from the provirus and converts the promoter into heterochromatin refractory to TNF-α induction. The present work represents the first demonstration of functional incompatibility between an otherwise functional NF-κB motif and a unique Sp1 site in the context of HIV-1 promoter. Our work provides important leads as per the evolution of HIV-1 subtype C viral promoter with relevance for gene expression regulation and viral latency.</p>',
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<div class="small-10 columns">
<h3>Epigenetic antibodies you can trust!</h3>
<p>Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level of siRNA knockdown validation to assure the specificity of our non-histone antibodies.</p>
<p><strong>Short interfering RNA (siRNA)</strong> degrades target mRNA, followed by the knock-down of protein production. If the antibody that recognizes the protein of interest is specific, the Western blot of siRNA-treated cells will show a significant reduction of signal vs. untreated cells.</p>
<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
<p class="text-center"><small>WB results obtained with the HDAC1 pAb (Cat. No. C15100144) <br />on siRNA transfected cells (lane 2) and on untransfected control cells (lane 1).</small></p>
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<p><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></p>
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<div class="spaced"></div>
<p style="text-align: left;"><span style="font-weight: 400;">The below list shows our first siRNA validated antibodies. More results - coming soon</span>.</p>',
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<h3>Epigenetic antibodies you can trust!</h3>
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<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
<p class="text-center"><small>WB results obtained with the HDAC1 pAb (Cat. No. C15100144) <br />on siRNA transfected cells (lane 2) and on untransfected control cells (lane 1).</small></p>
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<p><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></p>
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<p style="text-align: left;"><span style="font-weight: 400;">The below list shows our first siRNA validated antibodies. More results - coming soon</span>.</p>',
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<h3>Epigenetic antibodies you can trust!</h3>
<p>Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level of siRNA knockdown validation to assure the specificity of our non-histone antibodies.</p>
<p><strong>Short interfering RNA (siRNA)</strong> degrades target mRNA, followed by the knock-down of protein production. If the antibody that recognizes the protein of interest is specific, the Western blot of siRNA-treated cells will show a significant reduction of signal vs. untreated cells.</p>
<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
<p class="text-center"><small>WB results obtained with the HDAC1 pAb (Cat. No. C15100144) <br />on siRNA transfected cells (lane 2) and on untransfected control cells (lane 1).</small></p>
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<p style="text-align: left;"><span style="font-weight: 400;">The below list shows our first siRNA validated antibodies. More results - coming soon</span>.</p>'
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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<p><span>Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5 of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<br />
<div class="small-9 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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>
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<p></p>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<th>References</th>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:3,000</td>
<td>Fig 3</td>
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<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 4, 5</td>
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<td>Immunofluorescence</td>
<td>1:500</td>
<td>Fig 6</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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'description' => '<p><span>Alternative names: <strong>POLR2A</strong>, <strong>RPB1</strong>, <strong>POLR2</strong>, <strong>RPOL2</strong></span></p>
<p><span>Monoclonal antibody raised in mouse against the <strong>YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5</strong> of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="spacer"></div>
<div class="spacer"></div>
<div class="row">
<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
</div>
<p></p>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="spacer"></div>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<p></p>
<div class="spacer"></div>
<div class="spacer"></div>
<div class="row">
<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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>
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<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<br />
<div class="small-9 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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>
<p></p>
<div class="row">
<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
</div>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><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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<div class="small-10 columns">
<h3>Epigenetic antibodies you can trust!</h3>
<p>Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level of siRNA knockdown validation to assure the specificity of our non-histone antibodies.</p>
<p><strong>Short interfering RNA (siRNA)</strong> degrades target mRNA, followed by the knock-down of protein production. If the antibody that recognizes the protein of interest is specific, the Western blot of siRNA-treated cells will show a significant reduction of signal vs. untreated cells.</p>
<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
<p class="text-center"><small>WB results obtained with the HDAC1 pAb (Cat. No. C15100144) <br />on siRNA transfected cells (lane 2) and on untransfected control cells (lane 1).</small></p>
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<p><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></p>
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<p style="text-align: left;"><span style="font-weight: 400;">The below list shows our first siRNA validated antibodies. More results - coming soon</span>.</p>',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
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<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 provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</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>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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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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<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
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<p><span>Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5 of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p><span>Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5 of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<td>Western Blotting</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 μg per IP.</small></p>',
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'name' => 'Pol II S5p Antibody',
'description' => '<p><span>Alternative names: <strong>POLR2A</strong>, <strong>RPB1</strong>, <strong>POLR2</strong>, <strong>RPOL2</strong></span></p>
<p><span>Monoclonal antibody raised in mouse against the <strong>YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5</strong> of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="spacer"></div>
<div class="spacer"></div>
<div class="row">
<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
</div>
<p></p>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<div class="spacer"></div>
<div class="spacer"></div>
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<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
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<div class="small-10 columns">
<h3>Epigenetic antibodies you can trust!</h3>
<p>Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level of siRNA knockdown validation to assure the specificity of our non-histone antibodies.</p>
<p><strong>Short interfering RNA (siRNA)</strong> degrades target mRNA, followed by the knock-down of protein production. If the antibody that recognizes the protein of interest is specific, the Western blot of siRNA-treated cells will show a significant reduction of signal vs. untreated cells.</p>
<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
<p class="text-center"><small>WB results obtained with the HDAC1 pAb (Cat. No. C15100144) <br />on siRNA transfected cells (lane 2) and on untransfected control cells (lane 1).</small></p>
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<div class="small-2 columns">
<p><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></p>
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<div class="spaced"></div>
<p style="text-align: left;"><span style="font-weight: 400;">The below list shows our first siRNA validated antibodies. More results - coming soon</span>.</p>',
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<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 provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</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>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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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>
<div class="small-12 medium-6 large-6 columns">
<p></p>
<p></p>
<p></p>
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<p></p>
<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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'name' => 'CDK7 Inhibition is Effective in all the Subtypes of Breast Cancer: Determinants of Response and Synergy with EGFR Inhibition.',
'authors' => 'McDermott MSJ, Sharko AC, Munie J, Kassler S, Melendez T, Lim CU, Broude EV',
'description' => '<p>CDK7, a transcriptional cyclin-dependent kinase, is emerging as a novel cancer target. Triple-negative breast cancers (TNBC) but not estrogen receptor-positive (ER+) breast cancers have been reported to be uniquely sensitive to the CDK7 inhibitor THZ1 due to the inhibition of a cluster of TNBC-specific genes. However, bioinformatic analysis indicates that CDK7 RNA expression is associated with negative prognosis in all the major subtypes of breast cancer. To further elucidate the effects of CDK7 inhibition in breast cancer, we profiled a panel of cell lines representing different breast cancer subtypes. THZ1 inhibited cell growth in all subtypes (TNBC, HER2+, ER+, and HER2+/ER+) with no apparent subtype selectivity. THZ1 inhibited CDK7 activity and induced G1 arrest and apoptosis in all the tested cell lines, but THZ1 sensitivity did not correlate with CDK7 inhibition or CDK7 expression levels. THZ1 sensitivity across the cell line panel did not correlate with TNBC-specific gene expression but it was found to correlate with the differential inhibition of three genes: CDKN1B, MYC and transcriptional coregulator CITED2. Response to THZ1 also correlated with basal CITED2 protein expression, a potential marker of CDK7 inhibitor sensitivity. Furthermore, all of the THZ1-inhibited genes examined were inducible by EGF but THZ1 prevented this induction. THZ1 had synergistic or additive effects when combined with the EGFR inhibitor erlotinib, with no outward selectivity for a particular subtype of breast cancer. These results suggest a potential broad utility for CDK7 inhibitors in breast cancer therapy and the potential for combining CDK7 and EGFR inhibitors.</p>',
'date' => '2020-03-06',
'pmid' => 'http://www.pubmed.gov/32155786',
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'description' => '<p>How transcription affects genome 3D organization is not well understood. We found that during influenza A (IAV) infection, rampant transcription rapidly reorganizes host cell chromatin interactions. These changes occur at the ends of highly transcribed genes, where global inhibition of transcription termination by IAV NS1 protein causes readthrough transcription for hundreds of kilobases. In these readthrough regions, elongating RNA polymerase II disrupts chromatin interactions by inducing cohesin displacement from CTCF sites, leading to locus decompaction. Readthrough transcription into heterochromatin regions switches them from the inert (B) to the permissive (A) chromatin compartment and enables transcription factor binding. Data from non-viral transcription stimuli show that transcription similarly affects cohesin-mediated chromatin contacts within gene bodies. Conversely, inhibition of transcription elongation allows cohesin to accumulate at previously transcribed intragenic CTCF sites and to mediate chromatin looping and compaction. Our data indicate that transcription elongation by RNA polymerase II remodels genome 3D architecture.</p>',
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'pmid' => 'http://www.pubmed.gov/30146161',
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'description' => '<p>Although many long non-coding RNAs (lncRNAs) are imprinted, their roles often remain unknown. The Dlk1-Dio3 domain expresses the lncRNA Meg3 and multiple microRNAs and small nucleolar RNAs (snoRNAs) on the maternal chromosome and constitutes an epigenetic model for development. The domain's Dlk1 (Delta-like-1) gene encodes a ligand that inhibits Notch1 signaling and regulates diverse developmental processes. Using a hybrid embryonic stem cell (ESC) system, we find that Dlk1 becomes imprinted during neural differentiation and that this involves transcriptional upregulation on the paternal chromosome. The maternal Dlk1 gene remains poised. Its protection against activation is controlled in cis by Meg3 expression and also requires the H3-Lys-27 methyltransferase Ezh2. Maternal Meg3 expression additionally protects against de novo DNA methylation at its promoter. We find that Meg3 lncRNA is partially retained in cis and overlaps the maternal Dlk1 in embryonic cells. Combined, our data evoke an imprinting model in which allelic lncRNA expression prevents gene activation in cis.</p>',
'date' => '2018-04-10',
'pmid' => 'http://www.pubmed.gov/29641995',
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'description' => '<p>The nuclear factor-κB (NFκB) family of <span class="highlight">transcription</span> factors has been implicated in inflammatory disorders, viral infections, and cancer. Most of the drugs that inhibit NFκB show significant side effects, possibly due to sustained NFκB suppression. Drugs affecting induced, but not basal, NFκB activity may have the potential to provide therapeutic benefit without associated toxicity. NFκB activation by stress-inducible cell cycle inhibitor p21 was shown to be mediated by a p21-stimulated <span class="highlight">transcription</span>-regulating kinase <span class="highlight">CDK8</span>. <span class="highlight">CDK8</span> and its paralog CDK19, associated with the transcriptional <span class="highlight">Mediator</span> complex, act as coregulators of several <span class="highlight">transcription</span> factors implicated in cancer; <span class="highlight">CDK8</span>/<span class="highlight">19</span> inhibitors are entering clinical development. Here we show that <span class="highlight">CDK8</span>/<span class="highlight">19</span> inhibition by different small-molecule kinase inhibitors or shRNAs suppresses the elongation of NFκB-induced <span class="highlight">transcription</span> when such <span class="highlight">transcription</span> is activated by p21-independent canonical inducers, such as TNFα. On NFκB activation, <span class="highlight">CDK8</span>/<span class="highlight">19</span> are corecruited with NFκB to the promoters of the responsive genes. Inhibition of <span class="highlight">CDK8</span>/<span class="highlight">19</span> kinase activity suppresses the RNA polymerase II C-terminal domain phosphorylation required for transcriptional elongation, in a gene-specific manner. Genes coregulated by <span class="highlight">CDK8</span>/<span class="highlight">19</span> and NFκB include <i>IL8</i>, <i>CXCL1</i>, and <i>CXCL2</i>, which encode tumor-promoting proinflammatory cytokines. Although it suppressed newly induced NFκB-driven <span class="highlight">transcription</span>, <span class="highlight">CDK8</span>/<span class="highlight">19</span> inhibition in most cases had no effect on the basal expression of NFκB-regulated genes or promoters; the same selective regulation of newly induced <span class="highlight">transcription</span> was observed with other <span class="highlight">transcription</span> signals potentiated by <span class="highlight">CDK8</span>/<span class="highlight">19</span>. This selective role of <span class="highlight">CDK8</span>/<span class="highlight">19</span> identifies these <span class="highlight">kinases</span> as mediators of transcriptional reprogramming, a key aspect of development and differentiation as well as pathological processes.</p>',
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'authors' => 'Verma A et al.',
'description' => '<p>Of the various genetic subtypes of HIV-1, HIV-2 and SIV, only in subtype C of HIV-1, a genetically variant NF-κB binding site is found at the core of the viral promoter in association with a subtype-specific Sp1III motif. How the subtype-associated variations in the core transcription factor binding sites (TFBS) influence gene expression from the viral promoter has not been examined previously. Using panels of infectious viral molecular clones, we demonstrate that subtype-specific NF-κB and Sp1III motifs have evolved for optimal gene expression, and neither of the motifs can be substituted by a corresponding TFBS variant.The variant NF-κB motif binds NF-κB with an affinity two-fold higher than that of the generic NF-κB site. Importantly, in the context of an infectious virus, the subtype-specific Sp1III motif demonstrates a profound loss of function in association with the generic NF-κB motif. An additional substitution of the Sp1III motif fully restores viral replication suggesting that the subtype C specific Sp1III has evolved to function with the variant, but not generic, NF-κB motif. A change of only two base pairs in the central NF-κB motif completely suppresses viral transcription from the provirus and converts the promoter into heterochromatin refractory to TNF-α induction. The present work represents the first demonstration of functional incompatibility between an otherwise functional NF-κB motif and a unique Sp1 site in the context of HIV-1 promoter. Our work provides important leads as per the evolution of HIV-1 subtype C viral promoter with relevance for gene expression regulation and viral latency.</p>',
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<p><span>Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5 of the repeat sequence. </span></p>',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<td>Fig 1, 2</td>
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<td>1:3,000</td>
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<p><span>Monoclonal antibody raised in mouse against the YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5 of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="spacer"></div>
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<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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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<br />
<div class="small-9 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
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<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>1-2 μg/ChIP</td>
<td>Fig 1, 2</td>
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<td>1:3,000</td>
<td>Fig 3</td>
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<td>Fig 4, 5</td>
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<p><span>Monoclonal antibody raised in mouse against the <strong>YSPTSPS repeat in the B1 subunit of RNA polymerase II, phosphorylated at Ser5</strong> of the repeat sequence. </span></p>',
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIP.png" alt="Pol II S5p Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) and optimized PCR primer pairs for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010051), using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 μg of antibody per ChIP experiment was analyzed. IgG (2 μg/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter and the coding region of the constitutively expressed GAPDH and ACTB genes, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div class="spacer"></div>
<div class="spacer"></div>
<div class="row">
<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-A.png" alt="Pol II S5p Antibody ChIP-seq Grade" style="display: block; margin-left: auto; margin-right: auto;" /><br /> B. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-B.png" alt="Pol II S5p Antibody for ChIP-seq" style="display: block; margin-left: auto; margin-right: auto;" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-C.png" alt="Pol II S5p Antibody for ChIP-seq assay " style="display: block; margin-left: auto; margin-right: auto;" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/C15200007_ChIPSeq-D.png" alt="Pol II S5p Antibody validated in ChIP-seq " style="display: block; margin-left: auto; margin-right: auto;" /></div>
</div>
<p></p>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />ChIP was performed on sheared chromatin from 1 million HeLaS3 cells using 1 μg of the Diagenode antibody against Pol II S5p (Cat. No. C15200007) as described above. The IP’d DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure 2 shows the peak distribution along the complete sequence and a 150 kb region of the X-chromosome (figure 2A and B, respectively), and in a two genomic regions surrounding the GAPDH and ACTB positive control genes (figure 2C and D).<br /><br /></small></p>
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<div class="small-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_ELISA.png" alt="Pol II S5p Antibody ELISA validation" style="display: block; border: 1px solid gray; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Cross reactivity of the Diagenode monoclonal antibody directed against Pol IIS5p </strong><br />To test the specificity an ELISA was performed using a serial dilution of the Diagenode monoclonal antibody against Pol IIS5p (Cat. No. C15200007). The wells were coated with peptides containing the unmodified C-terminal repeat sequence as well as different phosphorylated peptides. Figure 3 shows the specificity of the antibody for the S5 phosphorylation.</small></p>
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<p></p>
<div class="spacer"></div>
<div class="spacer"></div>
<div class="row">
<div class="small-3 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-9 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Nuclear extracts (25 μg) from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against Pol II S5p (Cat. No. C15200007) 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>
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<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200007_WB_2.png" alt="Pol II S5p Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<br />
<div class="small-9 columns">
<p><small><strong>Figure 5. Western blot analysis using the Diagenode monoclonal antibody directed against Pol II S5p </strong><br />Whole cell extracts (40 μg) from HeLa cells transfected with Pol II siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using the Diagenode antibody against Pol II S5p (Cat. No. C15200007) 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"><img src="https://www.diagenode.com/img/product/antibodies/C15200007_IF.png" alt="Pol II S5p Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></div>
<div class="small-8 columns">
<p><small><strong>Figure 6. Immunofluorescence using the Diagenode monoclonal antibody directed against Pol II S5p</strong> <br />HeLa cells were stained with the Diagenode antibody against Pol II S5p (Cat. No. C15200007) and with DAPI. Cells were fixed with methanol and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the Pol II S5p antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<div class="small-10 columns">
<h3>Epigenetic antibodies you can trust!</h3>
<p>Antibody quality is essential for assay success. Diagenode offers antibodies that are actually validated and have been widely used and published by the scientific community. Now we are adding a new level of siRNA knockdown validation to assure the specificity of our non-histone antibodies.</p>
<p><strong>Short interfering RNA (siRNA)</strong> degrades target mRNA, followed by the knock-down of protein production. If the antibody that recognizes the protein of interest is specific, the Western blot of siRNA-treated cells will show a significant reduction of signal vs. untreated cells.</p>
<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
<p class="text-center"><small>WB results obtained with the HDAC1 pAb (Cat. No. C15100144) <br />on siRNA transfected cells (lane 2) and on untransfected control cells (lane 1).</small></p>
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<p><img src="https://www.diagenode.com/emailing/images/epi-success-guaranteed-icon.png" alt="Epigenetic success guaranteed" /></p>
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<p style="text-align: left;"><span style="font-weight: 400;">The below list shows our first siRNA validated antibodies. More results - coming soon</span>.</p>',
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<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 provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</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>CDK7, a transcriptional cyclin-dependent kinase, is emerging as a novel cancer target. Triple-negative breast cancers (TNBC) but not estrogen receptor-positive (ER+) breast cancers have been reported to be uniquely sensitive to the CDK7 inhibitor THZ1 due to the inhibition of a cluster of TNBC-specific genes. However, bioinformatic analysis indicates that CDK7 RNA expression is associated with negative prognosis in all the major subtypes of breast cancer. To further elucidate the effects of CDK7 inhibition in breast cancer, we profiled a panel of cell lines representing different breast cancer subtypes. THZ1 inhibited cell growth in all subtypes (TNBC, HER2+, ER+, and HER2+/ER+) with no apparent subtype selectivity. THZ1 inhibited CDK7 activity and induced G1 arrest and apoptosis in all the tested cell lines, but THZ1 sensitivity did not correlate with CDK7 inhibition or CDK7 expression levels. THZ1 sensitivity across the cell line panel did not correlate with TNBC-specific gene expression but it was found to correlate with the differential inhibition of three genes: CDKN1B, MYC and transcriptional coregulator CITED2. Response to THZ1 also correlated with basal CITED2 protein expression, a potential marker of CDK7 inhibitor sensitivity. Furthermore, all of the THZ1-inhibited genes examined were inducible by EGF but THZ1 prevented this induction. THZ1 had synergistic or additive effects when combined with the EGFR inhibitor erlotinib, with no outward selectivity for a particular subtype of breast cancer. These results suggest a potential broad utility for CDK7 inhibitors in breast cancer therapy and the potential for combining CDK7 and EGFR inhibitors.</p>',
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'description' => '<p>How transcription affects genome 3D organization is not well understood. We found that during influenza A (IAV) infection, rampant transcription rapidly reorganizes host cell chromatin interactions. These changes occur at the ends of highly transcribed genes, where global inhibition of transcription termination by IAV NS1 protein causes readthrough transcription for hundreds of kilobases. In these readthrough regions, elongating RNA polymerase II disrupts chromatin interactions by inducing cohesin displacement from CTCF sites, leading to locus decompaction. Readthrough transcription into heterochromatin regions switches them from the inert (B) to the permissive (A) chromatin compartment and enables transcription factor binding. Data from non-viral transcription stimuli show that transcription similarly affects cohesin-mediated chromatin contacts within gene bodies. Conversely, inhibition of transcription elongation allows cohesin to accumulate at previously transcribed intragenic CTCF sites and to mediate chromatin looping and compaction. Our data indicate that transcription elongation by RNA polymerase II remodels genome 3D architecture.</p>',
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'description' => '<p>Although many long non-coding RNAs (lncRNAs) are imprinted, their roles often remain unknown. The Dlk1-Dio3 domain expresses the lncRNA Meg3 and multiple microRNAs and small nucleolar RNAs (snoRNAs) on the maternal chromosome and constitutes an epigenetic model for development. The domain's Dlk1 (Delta-like-1) gene encodes a ligand that inhibits Notch1 signaling and regulates diverse developmental processes. Using a hybrid embryonic stem cell (ESC) system, we find that Dlk1 becomes imprinted during neural differentiation and that this involves transcriptional upregulation on the paternal chromosome. The maternal Dlk1 gene remains poised. Its protection against activation is controlled in cis by Meg3 expression and also requires the H3-Lys-27 methyltransferase Ezh2. Maternal Meg3 expression additionally protects against de novo DNA methylation at its promoter. We find that Meg3 lncRNA is partially retained in cis and overlaps the maternal Dlk1 in embryonic cells. Combined, our data evoke an imprinting model in which allelic lncRNA expression prevents gene activation in cis.</p>',
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<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
<p class="text-center"><small>WB results obtained with the HDAC1 pAb (Cat. No. C15100144) <br />on siRNA transfected cells (lane 2) and on untransfected control cells (lane 1).</small></p>
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<center><img src="https://www.diagenode.com/emailing/images/C15100144-wb.png" alt="" /></center>
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)
$externalLink = ' <a href="http://www.ncbi.nlm.nih.gov/pubmed/27194770" 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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