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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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/c15410337-chipseq-a.png" alt="BRD4 Antibody ChIP-seq Grade" /> <br />B. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-b.png" alt="BRD4 Antibody for ChIP-seq" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-c.png" alt="BRD4 Antibody for ChIP-seq assay" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-d.png" alt="BRD4 Antibody validated in ChIP-seq" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-elisa.jpg" alt="BRD4 Antibody ELISA Validation" /></p>
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<p><small><strong>Figure 3. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-wb.jpg" alt="BRD4 Antibody validated in Western Blot " /></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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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<td>2 µg/ChIP</td>
<td>Fig 1, 2</td>
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<td>1:10,000</td>
<td>Fig 3</td>
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<td>Fig 4</td>
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<p><small>*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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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-chip.jpg" alt="BRD4 Antibody ChIP Grade" /></p>
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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/c15410337-chipseq-a.png" alt="BRD4 Antibody ChIP-seq Grade" /> <br />B. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-b.png" alt="BRD4 Antibody for ChIP-seq" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-c.png" alt="BRD4 Antibody for ChIP-seq assay" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-d.png" alt="BRD4 Antibody validated in ChIP-seq" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-elisa.jpg" alt="BRD4 Antibody ELISA Validation" /></p>
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<p><small><strong>Figure 3. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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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<th>References</th>
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<td>ChIP/ChIP-seq *</td>
<td>2 µg/ChIP</td>
<td>Fig 1, 2</td>
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<tr>
<td>ELISA</td>
<td>1:10,000</td>
<td>Fig 3</td>
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<td>Western blotting</td>
<td>1:1,000</td>
<td>Fig 4</td>
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<p><small>*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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<div class="small-4 columns">
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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/c15410337-chipseq-a.png" alt="BRD4 Antibody ChIP-seq Grade" /> <br />B. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-b.png" alt="BRD4 Antibody for ChIP-seq" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-c.png" alt="BRD4 Antibody for ChIP-seq assay" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-d.png" alt="BRD4 Antibody validated in ChIP-seq" /></div>
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<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
</div>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-elisa.jpg" alt="BRD4 Antibody ELISA Validation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.</small></p>
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<div class="row">
<div class="small-2 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-wb.jpg" alt="BRD4 Antibody validated in Western Blot " /></p>
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<div class="small-10 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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="large-12 columns">Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on a genome-wide scale. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</div>
<div class="large-12 columns"></div>
<h5 class="large-12 columns"><strong></strong></h5>
<h5 class="large-12 columns"><strong>The ChIP-seq workflow</strong></h5>
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<div class="large-12 columns"><br />
<ol>
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<li class="large-12 columns"><strong>NGS Library Preparation</strong>: Ligate adapters and amplify IP'd material</li>
<li class="large-12 columns"><strong>Bioinformatic analysis</strong>: Perform r<span style="font-weight: 400;">ead filtering and trimming</span>, r<span style="font-weight: 400;">ead specific alignment, enrichment specific peak calling, QC metrics, multi-sample cross-comparison etc. </span></li>
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<div class="small-12 medium-10 large-9 small-centered columns">
<div class="radius panel" style="background-color: #fff;">
<h3 class="text-center" style="color: #b21329;">Need guidance?</h3>
<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
<div class="row">
<div class="small-6 medium-6 large-6 columns"><a href="../pages/which-kit-to-choose"><img alt="" src="https://www.diagenode.com/img/banners/banner-decide.png" /></a></div>
<div class="small-6 medium-6 large-6 columns"><a href="../pages/chip-kit-customizer-1"><img alt="" src="https://www.diagenode.com/img/banners/banner-customizer.png" /></a></div>
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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><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><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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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'info2' => '<p>BRD4 (UniProt/Swiss-Prot entry O60885) is a chromatin reader protein that binds acetylated histones. It remains associated with acetylated chromatin throughout the entire cell cycle and provides epigenetic memory for gene transcription by preserving an acetylated chromatin status. As such, it plays a key role in the transmission of epigenetic memory across cell divisions. BRD4 promotes phosphorylation of Ser-2 of the C-terminal domain (CTD) of RNA polymerase II and plays a key role in regulating the transcription of signal-inducible genes. It has been implicated in a translocation of chromosome 19 which causes an upper respiratory tract carcinoma.</p>',
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'slug' => 'brd4-polyclonal-antibody',
'meta_title' => 'BRD4 polyclonal antibody | Diagenode',
'meta_keywords' => 'BRD4 polyclonal antibody,Western blot,chromatin immunoprecipitation,Diagenode',
'meta_description' => ' BRD4 polyclonal antibody from diagenode raised in rabbit against human BRD4 (Bromodomain Containing 4), using two KLH-conjugated synthetic peptides from the N-terminal and central part of the protein, respectively has been validated for use in Western blot',
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'description' => 'BRD4 (UniProt/Swiss-Prot entry O60885) is a chromatin reader protein that binds acetylated histones. It remains associated with acetylated chromatin throughout the entire cell cycle and provides epigenetic memory for gene transcription by preserving an acetylated chromatin status. As such, it plays a key role in the transmission of epigenetic memory across cell divisions. BRD4 promotes phosphorylation of Ser-2 of the C-terminal domain (CTD) of RNA polymerase II and plays a key role in regulating the transcription of signal-inducible genes. It has been implicated in a translocation of chromosome 19 which causes an upper respiratory tract carcinoma.',
'clonality' => '',
'isotype' => 'NA',
'lot' => 'A2710P',
'concentration' => '2.6 µg/µl',
'reactivity' => 'Human',
'type' => 'Polyclonal',
'purity' => 'Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.',
'classification' => 'Classic',
'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution/amount</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP/ChIP-seq *</td>
<td>2 µg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:10,000</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Western blotting</td>
<td>1:1,000</td>
<td>Fig 4</td>
</tr>
</tbody>
</table>
<p></p>
<p><small>*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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'precautions' => 'This product is for research use only. Not for use in diagnostic or therapeutic procedures.',
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'name' => 'BRD4 Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against human <strong>BRD4</strong> <strong>(Bromodomain Containing 4)</strong>, using two KLH-conjugated synthetic peptides from the N-terminal and central part of the protein, respectively.</p>',
'label1' => 'Validation Data',
'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-chip.jpg" alt="BRD4 Antibody ChIP Grade" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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>
</div>
</div>
<div class="row">
<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-a.png" alt="BRD4 Antibody ChIP-seq Grade" /> <br />B. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-b.png" alt="BRD4 Antibody for ChIP-seq" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-c.png" alt="BRD4 Antibody for ChIP-seq assay" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-d.png" alt="BRD4 Antibody validated in ChIP-seq" /></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-elisa.jpg" alt="BRD4 Antibody ELISA Validation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-2 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-wb.jpg" alt="BRD4 Antibody validated in Western Blot " /></p>
</div>
<div class="small-10 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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="large-12 columns">Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on a genome-wide scale. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</div>
<div class="large-12 columns"></div>
<h5 class="large-12 columns"><strong></strong></h5>
<h5 class="large-12 columns"><strong>The ChIP-seq workflow</strong></h5>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/chip-seq-diagram.png" /></div>
<div class="large-12 columns"><br />
<ol>
<li class="large-12 columns"><strong>Chromatin preparation: </strong>Crosslink chromatin-bound proteins (histones or transcription factors) to DNA followed by cell lysis.</li>
<li class="large-12 columns"><strong>Chromatin shearing:</strong> Fragment chromatin by sonication to desired fragment size (100-500 bp)</li>
<li class="large-12 columns"><strong>Chromatin IP</strong>: Capture protein-DNA complexes with <strong><a href="../categories/chip-seq-grade-antibodies">specific ChIP-seq grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: Reverse cross-links, elute, and purify </li>
<li class="large-12 columns"><strong>NGS Library Preparation</strong>: Ligate adapters and amplify IP'd material</li>
<li class="large-12 columns"><strong>Bioinformatic analysis</strong>: Perform r<span style="font-weight: 400;">ead filtering and trimming</span>, r<span style="font-weight: 400;">ead specific alignment, enrichment specific peak calling, QC metrics, multi-sample cross-comparison etc. </span></li>
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</div>
</div>
<div class="row" style="margin-top: 32px;">
<div class="small-12 medium-10 large-9 small-centered columns">
<div class="radius panel" style="background-color: #fff;">
<h3 class="text-center" style="color: #b21329;">Need guidance?</h3>
<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
<div class="row">
<div class="small-6 medium-6 large-6 columns"><a href="../pages/which-kit-to-choose"><img alt="" src="https://www.diagenode.com/img/banners/banner-decide.png" /></a></div>
<div class="small-6 medium-6 large-6 columns"><a href="../pages/chip-kit-customizer-1"><img alt="" src="https://www.diagenode.com/img/banners/banner-customizer.png" /></a></div>
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'meta_title' => 'Chromatin Immunoprecipitation - ChIP-seq Kits - Dna methylation | Diagenode',
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<div class="small-12 medium-12 large-12 columns">Enzyme-linked immunosorbent assay.</div>
</div>',
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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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'description' => '<p><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<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>
<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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<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>
</div>
</div>
<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>
<div class="row">
<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>
</div>
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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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<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>
</div>
</div>
<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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'meta_description' => ' BRD4 polyclonal antibody from diagenode raised in rabbit against human BRD4 (Bromodomain Containing 4), using two KLH-conjugated synthetic peptides from the N-terminal and central part of the protein, respectively has been validated for use in Western blot',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-chip.jpg" alt="BRD4 Antibody ChIP Grade" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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="row">
<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-a.png" alt="BRD4 Antibody ChIP-seq Grade" /> <br />B. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-b.png" alt="BRD4 Antibody for ChIP-seq" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-c.png" alt="BRD4 Antibody for ChIP-seq assay" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-d.png" alt="BRD4 Antibody validated in ChIP-seq" /></div>
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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<div class="small-8 columns">
<p><small><strong>Figure 3. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<p><small><strong>Figure 3. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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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<td>2 µg/ChIP</td>
<td>Fig 1, 2</td>
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<td>1:10,000</td>
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<p><small>*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><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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/c15410337-chipseq-a.png" alt="BRD4 Antibody ChIP-seq Grade" /> <br />B. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-b.png" alt="BRD4 Antibody for ChIP-seq" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-c.png" alt="BRD4 Antibody for ChIP-seq assay" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-d.png" alt="BRD4 Antibody validated in ChIP-seq" /></div>
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<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-elisa.jpg" alt="BRD4 Antibody ELISA Validation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-2 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-wb.jpg" alt="BRD4 Antibody validated in Western Blot " /></p>
</div>
<div class="small-10 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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="large-12 columns">Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on a genome-wide scale. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</div>
<div class="large-12 columns"></div>
<h5 class="large-12 columns"><strong></strong></h5>
<h5 class="large-12 columns"><strong>The ChIP-seq workflow</strong></h5>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/chip-seq-diagram.png" /></div>
<div class="large-12 columns"><br />
<ol>
<li class="large-12 columns"><strong>Chromatin preparation: </strong>Crosslink chromatin-bound proteins (histones or transcription factors) to DNA followed by cell lysis.</li>
<li class="large-12 columns"><strong>Chromatin shearing:</strong> Fragment chromatin by sonication to desired fragment size (100-500 bp)</li>
<li class="large-12 columns"><strong>Chromatin IP</strong>: Capture protein-DNA complexes with <strong><a href="../categories/chip-seq-grade-antibodies">specific ChIP-seq grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: Reverse cross-links, elute, and purify </li>
<li class="large-12 columns"><strong>NGS Library Preparation</strong>: Ligate adapters and amplify IP'd material</li>
<li class="large-12 columns"><strong>Bioinformatic analysis</strong>: Perform r<span style="font-weight: 400;">ead filtering and trimming</span>, r<span style="font-weight: 400;">ead specific alignment, enrichment specific peak calling, QC metrics, multi-sample cross-comparison etc. </span></li>
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</div>
</div>
<div class="row" style="margin-top: 32px;">
<div class="small-12 medium-10 large-9 small-centered columns">
<div class="radius panel" style="background-color: #fff;">
<h3 class="text-center" style="color: #b21329;">Need guidance?</h3>
<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
<div class="row">
<div class="small-6 medium-6 large-6 columns"><a href="../pages/which-kit-to-choose"><img alt="" src="https://www.diagenode.com/img/banners/banner-decide.png" /></a></div>
<div class="small-6 medium-6 large-6 columns"><a href="../pages/chip-kit-customizer-1"><img alt="" src="https://www.diagenode.com/img/banners/banner-customizer.png" /></a></div>
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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><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>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>Learn more about: <a href="../applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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'description' => '<p>Molecular subtyping of cancer offers tremendous promise for the optimization of a precision oncology approach to anticancer therapy. Recent advances in pancreatic cancer research uncovered various molecular subtypes with tumors expressing a squamous/basal-like gene expression signature displaying a worse prognosis. Through unbiased epigenome mapping, we identified deltaNp63 as a major driver of a gene signature in pancreatic cancer cell lines, which we report to faithfully represent the highly aggressive pancreatic squamous subtype observed in vivo, and display the specific epigenetic marking of genes associated with decreased survival. Importantly, depletion of deltaNp63 in these systems significantly decreased cell proliferation and gene expression patterns associated with a squamous subtype and transcriptionally mimicked a subtype switch. Using genomic localization data of deltaNp63 in pancreatic cancer cell lines coupled with epigenome mapping data from patient-derived xenografts, we uncovered that deltaNp63 mainly exerts its effects by activating subtype-specific super enhancers. Furthermore, we identified a group of 45 subtype-specific super enhancers that are associated with poorer prognosis and are highly dependent on deltaNp63. Genes associated with these enhancers included a network of transcription factors, including HIF1A, BHLHE40, and RXRA, which form a highly intertwined transcriptional regulatory network with deltaNp63 to further activate downstream genes associated with poor survival.</p>',
'date' => '2018-12-26',
'pmid' => 'http://www.pubmed.gov/30541891',
'doi' => '10.1073/pnas.1812915116',
'modified' => '2019-06-07 09:29:25',
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<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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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<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
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<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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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'name' => 'BRD4 Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against human <strong>BRD4</strong> <strong>(Bromodomain Containing 4)</strong>, using two KLH-conjugated synthetic peptides from the N-terminal and central part of the protein, respectively.</p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-chip.jpg" alt="BRD4 Antibody ChIP Grade" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4</strong><br />ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and 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>
<div class="row">
<div class="small-12 columns">A. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-a.png" alt="BRD4 Antibody ChIP-seq Grade" /> <br />B. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-b.png" alt="BRD4 Antibody for ChIP-seq" /><br /> C. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-c.png" alt="BRD4 Antibody for ChIP-seq assay" /><br /> D. <img src="https://www.diagenode.com/img/product/antibodies/c15410337-chipseq-d.png" alt="BRD4 Antibody validated in ChIP-seq" /></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4</strong><br /> ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).</small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-elisa.jpg" alt="BRD4 Antibody ELISA Validation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Determination of the antibody titer</strong><br />To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.</small></p>
</div>
</div>
<div class="row">
<div class="small-2 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/c15410337-wb.jpg" alt="BRD4 Antibody validated in Western Blot " /></p>
</div>
<div class="small-10 columns">
<p><small><strong>Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4</strong><br />Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) 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>',
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'info2' => '<p>BRD4 (UniProt/Swiss-Prot entry O60885) is a chromatin reader protein that binds acetylated histones. It remains associated with acetylated chromatin throughout the entire cell cycle and provides epigenetic memory for gene transcription by preserving an acetylated chromatin status. As such, it plays a key role in the transmission of epigenetic memory across cell divisions. BRD4 promotes phosphorylation of Ser-2 of the C-terminal domain (CTD) of RNA polymerase II and plays a key role in regulating the transcription of signal-inducible genes. It has been implicated in a translocation of chromosome 19 which causes an upper respiratory tract carcinoma.</p>',
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<div class="large-12 columns">Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on a genome-wide scale. This technique is now used in a variety of life science disciplines including cellular differentiation, tumor suppressor gene silencing, and the effect of histone modifications on gene expression.</div>
<div class="large-12 columns"></div>
<h5 class="large-12 columns"><strong></strong></h5>
<h5 class="large-12 columns"><strong>The ChIP-seq workflow</strong></h5>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/chip-seq-diagram.png" /></div>
<div class="large-12 columns"><br />
<ol>
<li class="large-12 columns"><strong>Chromatin preparation: </strong>Crosslink chromatin-bound proteins (histones or transcription factors) to DNA followed by cell lysis.</li>
<li class="large-12 columns"><strong>Chromatin shearing:</strong> Fragment chromatin by sonication to desired fragment size (100-500 bp)</li>
<li class="large-12 columns"><strong>Chromatin IP</strong>: Capture protein-DNA complexes with <strong><a href="../categories/chip-seq-grade-antibodies">specific ChIP-seq grade antibodies</a></strong> against the histone or transcription factor of interest</li>
<li class="large-12 columns"><strong>DNA purification</strong>: Reverse cross-links, elute, and purify </li>
<li class="large-12 columns"><strong>NGS Library Preparation</strong>: Ligate adapters and amplify IP'd material</li>
<li class="large-12 columns"><strong>Bioinformatic analysis</strong>: Perform r<span style="font-weight: 400;">ead filtering and trimming</span>, r<span style="font-weight: 400;">ead specific alignment, enrichment specific peak calling, QC metrics, multi-sample cross-comparison etc. </span></li>
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<div class="small-12 medium-10 large-9 small-centered columns">
<div class="radius panel" style="background-color: #fff;">
<h3 class="text-center" style="color: #b21329;">Need guidance?</h3>
<p class="text-justify">Choose our full ChIP kits or simply choose what you need from antibodies, buffers, beads, chromatin shearing and purification reagents. With the ChIP Kit Customizer, you have complete flexibility on which components you want from our validated ChIP kits.</p>
<div class="row">
<div class="small-6 medium-6 large-6 columns"><a href="../pages/which-kit-to-choose"><img alt="" src="https://www.diagenode.com/img/banners/banner-decide.png" /></a></div>
<div class="small-6 medium-6 large-6 columns"><a href="../pages/chip-kit-customizer-1"><img alt="" src="https://www.diagenode.com/img/banners/banner-customizer.png" /></a></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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'description' => '<p><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<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>
<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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<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>
</div>
</div>
<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>
<div class="row">
<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>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><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>
</div>
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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>
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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>Learn more about: <a href="../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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$externalLink = ' <a href="http://www.pubmed.gov/30541891" 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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