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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2. </small></p>
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<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2. </small></p>
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<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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<p><strong>Figure 1. ChIP using the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing nuclease dead Cas9 and sgRNA targeting a sequence in intron 8 of the GAPDH gene, using the iDeal ChIP-seq kit for transcription factors. A titration consisting of 1, 2, 5 and 10 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) was tested. IgG (2 µg/IP) was used as negative IP control. qPCR was performed with primers specific for the targeted sequence in the GAPDH gene, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-a.jpg" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-b.jpg" width="700" /></center></div>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing dCas9 and a GAPDH sgRNA cells using 2 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 ChIP-seq profile in a region of chromosome 12 surrounding the GAPDH gene (fig 2B) and in a region of chromosome 2 surrounding an off-target peak in the YIPF4 gene.</p>
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<p><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells transfected with dCas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:5,000. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.</p>
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<p><strong>Figure 4. IP using the Diagenode antibody directed against Cas9</strong><br />IP was performed on whole cell extracts (500 µg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 µg) is shown in lane 1 and 2.</p>
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<p><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against Cas9</strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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 antibody (cat. No. C15310258) diluted 1:1000, followed by incubation with a goat anti-rabbit secondary antibody coupled to AF594. Nuclei were counter-stained with Hoechst 33342. Figure 5 shows the result in the presence (left) or absence (right) of doxycycline.</p>
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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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>
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'description' => '<p>Diagenode offers the broad range of antibodies raised against the N- or C-terminus of the Cas9 nuclease from <em>Streptococcus <g class="gr_ gr_5 gr-alert gr_spell gr_disable_anim_appear ContextualSpelling ins-del multiReplace" id="5" data-gr-id="5">pyogenes</g></em>. These highly specific polyclonal and monoclonal antibodies are validated in Western blot, immunoprecipitation, immunofluorescence and in chromatin immunoprecipitation.</p>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h2><em><a name="pyogenes"></a>S. pyogenes</em> CRISPR/Cas9 antibodies<a></a></h2>
<div class="panel">
<h2>Discover our first monoclonal CRISPR/Cas9 antibody validated in ChIP<br /><br /></h2>
<div class="row">
<div class="small-5 medium-5 large-5 columns"><img src="/img/landing-pages/crispr-cas9-chip-on-hih3t3.jpg" alt="" /></div>
<div class="small-7 medium-7 large-7 columns">
<ul>
<li>Validated in chromatin immunoprecipitation</li>
<li>Performs better than FLAG antibody</li>
<li>Excellent for WB, IF and IP</li>
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<p><small><strong>ChIP</strong> was performed on NIH3T3 cells stably expressing GFP-H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 µg chromatin was incubated overnight at 4°C with 5 or 10 µg of either an anti-FLAG antibody or the Diagenode antibody against Cas9 (Cat. No. C15200229). Mouse IgG was used as a negative IP control. qPCR was performed with primers specific for the GFP gene, and for a non-targeted region (protein kinase C delta, Prkcd), used as negative control. 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>
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<div class="row">
<div class="small-12 medium-12 large-12 columns text-right"><a href="/p/crispr-cas9-monoclonal-antibody-50-ug-25-μl" class="tiny details button radius">Learn more</a></div>
</div>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>First ChIP-grade CRISPR/Cas9 polyclonal antibody</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/landing-pages/c_a_s9-chip-grade-antibody.png" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Excellent polyclonal antibody for chromatin immunoprecipitation</li>
<li>Optimized for highest ChIP specificity and yields</li>
<li>Validated for all applications including immunoblotting, immunofluorescence and western blot</li>
</ul>
<p><small><strong>ChIP</strong> was performed on NIH3T3 cells stably expressing GFP- H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 μg chromatin was incubated with either 5 μg of an anti-FLAG antibody or 2 μl of the Diagenode antibody against Cas9. The pre-immune serum (PPI) was used as negative IP control. Then qPCR was performed with primers specific for the GFP gene, and for two non-targeted regions: Ppap2c and Prkcd, used as negative controls. This figure shows the recovery, expressed as a % of input.</small></p>
<p class="text-right"><a href="../p/crispr-cas9-polyclonal-antibody" class="details tiny button">Learn more</a></p>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>CRISPR/Cas9 monoclonal antibody 4G10</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/landing-pages/cas9_4g10_fig1.png" width="170" height="302" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Antibody raised against N-terminus of Cas9 nuclease</li>
<li>Validated for western blot, IP and immunofluorescence</li>
</ul>
<p><small><strong>Immunofluorescence</strong>: Hela cells were transiently transfected with a Cas9 expression vector. The cells were fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA. The cells were stained with the Cas9 antibody at 4°C o/n, followed by incubation with an anti mouse secondary antibody coupled to AF488 for 1 h at RT. Nuclei were counter-stained with Hoechst 33342 (right).</small></p>
<p class="text-right"><a href="../p/crispr-cas9-monoclonal-antibody-4g10-50-ug" class="details tiny button">Learn more</a></p>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>CRISPR/Cas9 C-terminal monoclonal antibody</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IP.png" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Antibody raised against C-terminus of Cas9 nuclease</li>
<li>Validated for western blot, IP and immunofluorescence</li>
</ul>
<p><small><strong> Western blot</strong> was performed on 20 μg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9. The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
<p class="text-right"><a href="../p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug" class="details tiny button">Learn more</a></p>
</div>
<div class="small-12 medium-12 large-12 columns">
<h3>Which CRISPR/Cas9 antibody is the best for your application?</h3>
<a name="table"></a>
<table>
<thead>
<tr>
<th>Antibody</th>
<th>WB</th>
<th>IF</th>
<th>IP</th>
<th>ChIP</th>
<th>Antibody raised against</th>
</tr>
</thead>
<tbody>
<tr>
<td><a href="../p/crispr-cas9-monoclonal-antibody-50-ug-25-μl"><strong class="diacol">CRISPR/Cas9 monoclonal antibody</strong></a></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><span class="diacol">N-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-polyclonal-antibody">CRISPR/Cas9 polyclonal antibody</a></td>
<td>++</td>
<td>++</td>
<td>++</td>
<td><strong class="diacol">+++</strong></td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-monoclonal-antibody-4g10-50-ug">CRISPR/Cas9 monoclonal antibody 4G10</a></td>
<td>+++</td>
<td>+++</td>
<td>++</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="../p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug-23-ul"><strong class="diacol">CRISPR/Cas9 C-terminal monoclonal antibody</strong></a> <span class="label alert" style="font-size: 0.9rem;">NEW!</span></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">no</strong></td>
<td><strong class="diacol">+</strong></td>
<td><span class="diacol">C-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug">CRISPR/Cas9 C-terminal monoclonal antibody</a></td>
<td>++</td>
<td>++</td>
<td>+</td>
<td>no</td>
<td><span class="diacol">C-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-monoclonal-antibody-7A9-50-mg">CRISPR/Cas9 monoclonal antibody 7A9</a></td>
<td>++</td>
<td>++</td>
<td>++</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-hrp-monoclonal-antibody-50-ul">CRISPR/Cas9 - HRP monoclonal antibody 7A9</a></td>
<td>+++</td>
<td>no</td>
<td>no</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
</tbody>
</table>
</div>
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'name' => 'CRISPR/Cas9 C-terminal Antibody',
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<p>Monoclonal antibody raised in mouse against the C-terminus of Cas9 nuclease (CRISPR-associated protein 9)</p>',
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<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-WB.png" alt="Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-WB2.png" alt="Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2. </small></p>
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<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 μg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 μg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 μg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 μg) is shown in lane 1 and 2. </small></p>
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<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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<a href="/jp/p/crispr-cas9-polyclonal-antibody"><img src="/img/product/antibodies/cas9-icon.png" alt="CRISPR/Cas9 Antibody " class="th"/></a> </div>
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<p><strong>Figure 1. ChIP using the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing nuclease dead Cas9 and sgRNA targeting a sequence in intron 8 of the GAPDH gene, using the iDeal ChIP-seq kit for transcription factors. A titration consisting of 1, 2, 5 and 10 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) was tested. IgG (2 µg/IP) was used as negative IP control. qPCR was performed with primers specific for the targeted sequence in the GAPDH gene, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-a.jpg" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-b.jpg" width="700" /></center></div>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing dCas9 and a GAPDH sgRNA cells using 2 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 ChIP-seq profile in a region of chromosome 12 surrounding the GAPDH gene (fig 2B) and in a region of chromosome 2 surrounding an off-target peak in the YIPF4 gene.</p>
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<p><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells transfected with dCas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:5,000. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.</p>
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<p><strong>Figure 4. IP using the Diagenode antibody directed against Cas9</strong><br />IP was performed on whole cell extracts (500 µg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 µg) is shown in lane 1 and 2.</p>
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<p><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against Cas9</strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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 antibody (cat. No. C15310258) diluted 1:1000, followed by incubation with a goat anti-rabbit secondary antibody coupled to AF594. Nuclei were counter-stained with Hoechst 33342. Figure 5 shows the result in the presence (left) or absence (right) of doxycycline.</p>
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 μg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 μg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 μg) is shown in lane 1 and 2. </small></p>
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<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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<p><strong>Figure 1. ChIP using the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing nuclease dead Cas9 and sgRNA targeting a sequence in intron 8 of the GAPDH gene, using the iDeal ChIP-seq kit for transcription factors. A titration consisting of 1, 2, 5 and 10 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) was tested. IgG (2 µg/IP) was used as negative IP control. qPCR was performed with primers specific for the targeted sequence in the GAPDH gene, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-a.jpg" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-b.jpg" width="700" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing dCas9 and a GAPDH sgRNA cells using 2 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 ChIP-seq profile in a region of chromosome 12 surrounding the GAPDH gene (fig 2B) and in a region of chromosome 2 surrounding an off-target peak in the YIPF4 gene.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-WB.png" /></center></div>
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<p><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells transfected with dCas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:5,000. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.</p>
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<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-IP.png" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 4. IP using the Diagenode antibody directed against Cas9</strong><br />IP was performed on whole cell extracts (500 µg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 µg) is shown in lane 1 and 2.</p>
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<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-IF.png" width="500" /></center></div>
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<div class="row">
<div class="small-12 columns">
<p><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against Cas9</strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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 antibody (cat. No. C15310258) diluted 1:1000, followed by incubation with a goat anti-rabbit secondary antibody coupled to AF594. Nuclei were counter-stained with Hoechst 33342. Figure 5 shows the result in the presence (left) or absence (right) of doxycycline.</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>
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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><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>
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'description' => '<p>Diagenode offers the broad range of antibodies raised against the N- or C-terminus of the Cas9 nuclease from <em>Streptococcus <g class="gr_ gr_5 gr-alert gr_spell gr_disable_anim_appear ContextualSpelling ins-del multiReplace" id="5" data-gr-id="5">pyogenes</g></em>. These highly specific polyclonal and monoclonal antibodies are validated in Western blot, immunoprecipitation, immunofluorescence and in chromatin immunoprecipitation.</p>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h2><em><a name="pyogenes"></a>S. pyogenes</em> CRISPR/Cas9 antibodies<a></a></h2>
<div class="panel">
<h2>Discover our first monoclonal CRISPR/Cas9 antibody validated in ChIP<br /><br /></h2>
<div class="row">
<div class="small-5 medium-5 large-5 columns"><img src="/img/landing-pages/crispr-cas9-chip-on-hih3t3.jpg" alt="" /></div>
<div class="small-7 medium-7 large-7 columns">
<ul>
<li>Validated in chromatin immunoprecipitation</li>
<li>Performs better than FLAG antibody</li>
<li>Excellent for WB, IF and IP</li>
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<p><small><strong>ChIP</strong> was performed on NIH3T3 cells stably expressing GFP-H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 µg chromatin was incubated overnight at 4°C with 5 or 10 µg of either an anti-FLAG antibody or the Diagenode antibody against Cas9 (Cat. No. C15200229). Mouse IgG was used as a negative IP control. qPCR was performed with primers specific for the GFP gene, and for a non-targeted region (protein kinase C delta, Prkcd), used as negative control. 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 medium-12 large-12 columns text-right"><a href="/p/crispr-cas9-monoclonal-antibody-50-ug-25-μl" class="tiny details button radius">Learn more</a></div>
</div>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>First ChIP-grade CRISPR/Cas9 polyclonal antibody</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/landing-pages/c_a_s9-chip-grade-antibody.png" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Excellent polyclonal antibody for chromatin immunoprecipitation</li>
<li>Optimized for highest ChIP specificity and yields</li>
<li>Validated for all applications including immunoblotting, immunofluorescence and western blot</li>
</ul>
<p><small><strong>ChIP</strong> was performed on NIH3T3 cells stably expressing GFP- H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 μg chromatin was incubated with either 5 μg of an anti-FLAG antibody or 2 μl of the Diagenode antibody against Cas9. The pre-immune serum (PPI) was used as negative IP control. Then qPCR was performed with primers specific for the GFP gene, and for two non-targeted regions: Ppap2c and Prkcd, used as negative controls. This figure shows the recovery, expressed as a % of input.</small></p>
<p class="text-right"><a href="../p/crispr-cas9-polyclonal-antibody" class="details tiny button">Learn more</a></p>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>CRISPR/Cas9 monoclonal antibody 4G10</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/landing-pages/cas9_4g10_fig1.png" width="170" height="302" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Antibody raised against N-terminus of Cas9 nuclease</li>
<li>Validated for western blot, IP and immunofluorescence</li>
</ul>
<p><small><strong>Immunofluorescence</strong>: Hela cells were transiently transfected with a Cas9 expression vector. The cells were fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA. The cells were stained with the Cas9 antibody at 4°C o/n, followed by incubation with an anti mouse secondary antibody coupled to AF488 for 1 h at RT. Nuclei were counter-stained with Hoechst 33342 (right).</small></p>
<p class="text-right"><a href="../p/crispr-cas9-monoclonal-antibody-4g10-50-ug" class="details tiny button">Learn more</a></p>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>CRISPR/Cas9 C-terminal monoclonal antibody</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IP.png" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Antibody raised against C-terminus of Cas9 nuclease</li>
<li>Validated for western blot, IP and immunofluorescence</li>
</ul>
<p><small><strong> Western blot</strong> was performed on 20 μg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9. The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
<p class="text-right"><a href="../p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug" class="details tiny button">Learn more</a></p>
</div>
<div class="small-12 medium-12 large-12 columns">
<h3>Which CRISPR/Cas9 antibody is the best for your application?</h3>
<a name="table"></a>
<table>
<thead>
<tr>
<th>Antibody</th>
<th>WB</th>
<th>IF</th>
<th>IP</th>
<th>ChIP</th>
<th>Antibody raised against</th>
</tr>
</thead>
<tbody>
<tr>
<td><a href="../p/crispr-cas9-monoclonal-antibody-50-ug-25-μl"><strong class="diacol">CRISPR/Cas9 monoclonal antibody</strong></a></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><span class="diacol">N-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-polyclonal-antibody">CRISPR/Cas9 polyclonal antibody</a></td>
<td>++</td>
<td>++</td>
<td>++</td>
<td><strong class="diacol">+++</strong></td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-monoclonal-antibody-4g10-50-ug">CRISPR/Cas9 monoclonal antibody 4G10</a></td>
<td>+++</td>
<td>+++</td>
<td>++</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="../p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug-23-ul"><strong class="diacol">CRISPR/Cas9 C-terminal monoclonal antibody</strong></a> <span class="label alert" style="font-size: 0.9rem;">NEW!</span></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">no</strong></td>
<td><strong class="diacol">+</strong></td>
<td><span class="diacol">C-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug">CRISPR/Cas9 C-terminal monoclonal antibody</a></td>
<td>++</td>
<td>++</td>
<td>+</td>
<td>no</td>
<td><span class="diacol">C-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-monoclonal-antibody-7A9-50-mg">CRISPR/Cas9 monoclonal antibody 7A9</a></td>
<td>++</td>
<td>++</td>
<td>++</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-hrp-monoclonal-antibody-50-ul">CRISPR/Cas9 - HRP monoclonal antibody 7A9</a></td>
<td>+++</td>
<td>no</td>
<td>no</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
</tbody>
</table>
</div>
</div>
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2. </small></p>
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<p><strong>Figure 1. ChIP using the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing nuclease dead Cas9 and sgRNA targeting a sequence in intron 8 of the GAPDH gene, using the iDeal ChIP-seq kit for transcription factors. A titration consisting of 1, 2, 5 and 10 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) was tested. IgG (2 µg/IP) was used as negative IP control. qPCR was performed with primers specific for the targeted sequence in the GAPDH gene, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing dCas9 and a GAPDH sgRNA cells using 2 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 ChIP-seq profile in a region of chromosome 12 surrounding the GAPDH gene (fig 2B) and in a region of chromosome 2 surrounding an off-target peak in the YIPF4 gene.</p>
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<p><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells transfected with dCas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:5,000. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.</p>
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<p><strong>Figure 4. IP using the Diagenode antibody directed against Cas9</strong><br />IP was performed on whole cell extracts (500 µg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 µg) is shown in lane 1 and 2.</p>
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-WB2.png" alt="Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
</div>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IP.png" alt="IP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2. </small></p>
</div>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
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<div class="extra-spaced"></div>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IF.png" alt="IP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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<p><strong>Figure 1. ChIP using the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing nuclease dead Cas9 and sgRNA targeting a sequence in intron 8 of the GAPDH gene, using the iDeal ChIP-seq kit for transcription factors. A titration consisting of 1, 2, 5 and 10 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) was tested. IgG (2 µg/IP) was used as negative IP control. qPCR was performed with primers specific for the targeted sequence in the GAPDH gene, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-a.jpg" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-b.jpg" width="700" /></center></div>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing dCas9 and a GAPDH sgRNA cells using 2 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 ChIP-seq profile in a region of chromosome 12 surrounding the GAPDH gene (fig 2B) and in a region of chromosome 2 surrounding an off-target peak in the YIPF4 gene.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-WB.png" /></center></div>
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<p><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells transfected with dCas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:5,000. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-IP.png" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 4. IP using the Diagenode antibody directed against Cas9</strong><br />IP was performed on whole cell extracts (500 µg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 µg) is shown in lane 1 and 2.</p>
</div>
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<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-IF.png" width="500" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against Cas9</strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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 antibody (cat. No. C15310258) diluted 1:1000, followed by incubation with a goat anti-rabbit secondary antibody coupled to AF594. Nuclei were counter-stained with Hoechst 33342. Figure 5 shows the result in the presence (left) or absence (right) of doxycycline.</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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<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><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>
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<li>Expert technical support</li>
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'description' => '<p>Diagenode offers the broad range of antibodies raised against the N- or C-terminus of the Cas9 nuclease from <em>Streptococcus <g class="gr_ gr_5 gr-alert gr_spell gr_disable_anim_appear ContextualSpelling ins-del multiReplace" id="5" data-gr-id="5">pyogenes</g></em>. These highly specific polyclonal and monoclonal antibodies are validated in Western blot, immunoprecipitation, immunofluorescence and in chromatin immunoprecipitation.</p>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h2><em><a name="pyogenes"></a>S. pyogenes</em> CRISPR/Cas9 antibodies<a></a></h2>
<div class="panel">
<h2>Discover our first monoclonal CRISPR/Cas9 antibody validated in ChIP<br /><br /></h2>
<div class="row">
<div class="small-5 medium-5 large-5 columns"><img src="/img/landing-pages/crispr-cas9-chip-on-hih3t3.jpg" alt="" /></div>
<div class="small-7 medium-7 large-7 columns">
<ul>
<li>Validated in chromatin immunoprecipitation</li>
<li>Performs better than FLAG antibody</li>
<li>Excellent for WB, IF and IP</li>
</ul>
<p><small><strong>ChIP</strong> was performed on NIH3T3 cells stably expressing GFP-H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 µg chromatin was incubated overnight at 4°C with 5 or 10 µg of either an anti-FLAG antibody or the Diagenode antibody against Cas9 (Cat. No. C15200229). Mouse IgG was used as a negative IP control. qPCR was performed with primers specific for the GFP gene, and for a non-targeted region (protein kinase C delta, Prkcd), used as negative control. 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 medium-12 large-12 columns text-right"><a href="/p/crispr-cas9-monoclonal-antibody-50-ug-25-μl" class="tiny details button radius">Learn more</a></div>
</div>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>First ChIP-grade CRISPR/Cas9 polyclonal antibody</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/landing-pages/c_a_s9-chip-grade-antibody.png" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Excellent polyclonal antibody for chromatin immunoprecipitation</li>
<li>Optimized for highest ChIP specificity and yields</li>
<li>Validated for all applications including immunoblotting, immunofluorescence and western blot</li>
</ul>
<p><small><strong>ChIP</strong> was performed on NIH3T3 cells stably expressing GFP- H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 μg chromatin was incubated with either 5 μg of an anti-FLAG antibody or 2 μl of the Diagenode antibody against Cas9. The pre-immune serum (PPI) was used as negative IP control. Then qPCR was performed with primers specific for the GFP gene, and for two non-targeted regions: Ppap2c and Prkcd, used as negative controls. This figure shows the recovery, expressed as a % of input.</small></p>
<p class="text-right"><a href="../p/crispr-cas9-polyclonal-antibody" class="details tiny button">Learn more</a></p>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>CRISPR/Cas9 monoclonal antibody 4G10</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/landing-pages/cas9_4g10_fig1.png" width="170" height="302" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Antibody raised against N-terminus of Cas9 nuclease</li>
<li>Validated for western blot, IP and immunofluorescence</li>
</ul>
<p><small><strong>Immunofluorescence</strong>: Hela cells were transiently transfected with a Cas9 expression vector. The cells were fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA. The cells were stained with the Cas9 antibody at 4°C o/n, followed by incubation with an anti mouse secondary antibody coupled to AF488 for 1 h at RT. Nuclei were counter-stained with Hoechst 33342 (right).</small></p>
<p class="text-right"><a href="../p/crispr-cas9-monoclonal-antibody-4g10-50-ug" class="details tiny button">Learn more</a></p>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>CRISPR/Cas9 C-terminal monoclonal antibody</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IP.png" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Antibody raised against C-terminus of Cas9 nuclease</li>
<li>Validated for western blot, IP and immunofluorescence</li>
</ul>
<p><small><strong> Western blot</strong> was performed on 20 μg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9. The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
<p class="text-right"><a href="../p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug" class="details tiny button">Learn more</a></p>
</div>
<div class="small-12 medium-12 large-12 columns">
<h3>Which CRISPR/Cas9 antibody is the best for your application?</h3>
<a name="table"></a>
<table>
<thead>
<tr>
<th>Antibody</th>
<th>WB</th>
<th>IF</th>
<th>IP</th>
<th>ChIP</th>
<th>Antibody raised against</th>
</tr>
</thead>
<tbody>
<tr>
<td><a href="../p/crispr-cas9-monoclonal-antibody-50-ug-25-μl"><strong class="diacol">CRISPR/Cas9 monoclonal antibody</strong></a></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><span class="diacol">N-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-polyclonal-antibody">CRISPR/Cas9 polyclonal antibody</a></td>
<td>++</td>
<td>++</td>
<td>++</td>
<td><strong class="diacol">+++</strong></td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-monoclonal-antibody-4g10-50-ug">CRISPR/Cas9 monoclonal antibody 4G10</a></td>
<td>+++</td>
<td>+++</td>
<td>++</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="../p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug-23-ul"><strong class="diacol">CRISPR/Cas9 C-terminal monoclonal antibody</strong></a> <span class="label alert" style="font-size: 0.9rem;">NEW!</span></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">no</strong></td>
<td><strong class="diacol">+</strong></td>
<td><span class="diacol">C-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug">CRISPR/Cas9 C-terminal monoclonal antibody</a></td>
<td>++</td>
<td>++</td>
<td>+</td>
<td>no</td>
<td><span class="diacol">C-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-monoclonal-antibody-7A9-50-mg">CRISPR/Cas9 monoclonal antibody 7A9</a></td>
<td>++</td>
<td>++</td>
<td>++</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-hrp-monoclonal-antibody-50-ul">CRISPR/Cas9 - HRP monoclonal antibody 7A9</a></td>
<td>+++</td>
<td>no</td>
<td>no</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
</tbody>
</table>
</div>
</div>
</div>',
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 μg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 μg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 μg) is shown in lane 1 and 2. </small></p>
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<span class="success label" style="">C15310258-100</span>
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<p><strong>Figure 1. ChIP using the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing nuclease dead Cas9 and sgRNA targeting a sequence in intron 8 of the GAPDH gene, using the iDeal ChIP-seq kit for transcription factors. A titration consisting of 1, 2, 5 and 10 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) was tested. IgG (2 µg/IP) was used as negative IP control. qPCR was performed with primers specific for the targeted sequence in the GAPDH gene, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing dCas9 and a GAPDH sgRNA cells using 2 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 ChIP-seq profile in a region of chromosome 12 surrounding the GAPDH gene (fig 2B) and in a region of chromosome 2 surrounding an off-target peak in the YIPF4 gene.</p>
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<p><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells transfected with dCas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:5,000. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.</p>
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<p><strong>Figure 4. IP using the Diagenode antibody directed against Cas9</strong><br />IP was performed on whole cell extracts (500 µg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 µg) is shown in lane 1 and 2.</p>
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<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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<td>Western Blotting</td>
<td>1:1,000 - 1:5,000</td>
<td>Fig 1,2</td>
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<td>Fig 3</td>
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<p>Monoclonal antibody raised in mouse against the C-terminus of Cas9 nuclease (CRISPR-associated protein 9)</p>',
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<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-WB.png" alt="Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-WB2.png" alt="Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IP.png" alt="IP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IF.png" alt="IP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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'name' => 'CRISPR/Cas9 Antibody',
'description' => '<p>Polyclonal antibody raised in rabbit against the <strong>Cas9</strong> nuclease (<strong>CRISPR</strong>-associated protein 9) using a recombinant protein. </p>',
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-fig1.png" /></center></div>
<div class="small-6 columns">
<p><strong>Figure 1. ChIP using the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing nuclease dead Cas9 and sgRNA targeting a sequence in intron 8 of the GAPDH gene, using the iDeal ChIP-seq kit for transcription factors. A titration consisting of 1, 2, 5 and 10 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) was tested. IgG (2 µg/IP) was used as negative IP control. qPCR was performed with primers specific for the targeted sequence in the GAPDH gene, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-a.jpg" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-b.jpg" width="700" /></center></div>
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<div class="small-12 columns">
<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing dCas9 and a GAPDH sgRNA cells using 2 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 ChIP-seq profile in a region of chromosome 12 surrounding the GAPDH gene (fig 2B) and in a region of chromosome 2 surrounding an off-target peak in the YIPF4 gene.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-WB.png" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells transfected with dCas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:5,000. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.</p>
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<div class="row">
<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-IP.png" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 4. IP using the Diagenode antibody directed against Cas9</strong><br />IP was performed on whole cell extracts (500 µg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 µg) is shown in lane 1 and 2.</p>
</div>
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<div class="row">
<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-IF.png" width="500" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against Cas9</strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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 antibody (cat. No. C15310258) diluted 1:1000, followed by incubation with a goat anti-rabbit secondary antibody coupled to AF594. Nuclei were counter-stained with Hoechst 33342. Figure 5 shows the result in the presence (left) or absence (right) of doxycycline.</p>
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'slug' => 'crispr-cas9-polyclonal-antibody',
'meta_title' => 'CRISPR/Cas9 Antibody - ChIP-seq Grade (C15310258) | Diagenode',
'meta_keywords' => 'Cas9, CRISPR, CAs9/CRISPR, crispr cas 9, crispr technology, gene editing crispr, genome engineering, genomic engineering, genome editing, crispr system, cas9 genome editing, targeted genome editing, crispr cas9 genome editing, cas9 nuclease, cas9 endonucl',
'meta_description' => 'S. pyogenes CRISPR/Cas9 Polyclonal Antibody validated in ChIP-seq, ChIP-qPCR, IP, IF and WB. Batch-specific data available on the website. Sample size available.',
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<p>Diagenode offers huge selection of highly sensitive antibodies validated in IF.</p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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'name' => '<em>S. pyogenes</EM> CRISPR/Cas9 antibodies',
'description' => '<p>Diagenode offers the broad range of antibodies raised against the N- or C-terminus of the Cas9 nuclease from <em>Streptococcus <g class="gr_ gr_5 gr-alert gr_spell gr_disable_anim_appear ContextualSpelling ins-del multiReplace" id="5" data-gr-id="5">pyogenes</g></em>. These highly specific polyclonal and monoclonal antibodies are validated in Western blot, immunoprecipitation, immunofluorescence and in chromatin immunoprecipitation.</p>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h2><em><a name="pyogenes"></a>S. pyogenes</em> CRISPR/Cas9 antibodies<a></a></h2>
<div class="panel">
<h2>Discover our first monoclonal CRISPR/Cas9 antibody validated in ChIP<br /><br /></h2>
<div class="row">
<div class="small-5 medium-5 large-5 columns"><img src="/img/landing-pages/crispr-cas9-chip-on-hih3t3.jpg" alt="" /></div>
<div class="small-7 medium-7 large-7 columns">
<ul>
<li>Validated in chromatin immunoprecipitation</li>
<li>Performs better than FLAG antibody</li>
<li>Excellent for WB, IF and IP</li>
</ul>
<p><small><strong>ChIP</strong> was performed on NIH3T3 cells stably expressing GFP-H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 µg chromatin was incubated overnight at 4°C with 5 or 10 µg of either an anti-FLAG antibody or the Diagenode antibody against Cas9 (Cat. No. C15200229). Mouse IgG was used as a negative IP control. qPCR was performed with primers specific for the GFP gene, and for a non-targeted region (protein kinase C delta, Prkcd), used as negative control. 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 medium-12 large-12 columns text-right"><a href="/p/crispr-cas9-monoclonal-antibody-50-ug-25-μl" class="tiny details button radius">Learn more</a></div>
</div>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>First ChIP-grade CRISPR/Cas9 polyclonal antibody</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/landing-pages/c_a_s9-chip-grade-antibody.png" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Excellent polyclonal antibody for chromatin immunoprecipitation</li>
<li>Optimized for highest ChIP specificity and yields</li>
<li>Validated for all applications including immunoblotting, immunofluorescence and western blot</li>
</ul>
<p><small><strong>ChIP</strong> was performed on NIH3T3 cells stably expressing GFP- H2B, nuclease dead Cas9, and a GFP-targeting gRNA. 50 μg chromatin was incubated with either 5 μg of an anti-FLAG antibody or 2 μl of the Diagenode antibody against Cas9. The pre-immune serum (PPI) was used as negative IP control. Then qPCR was performed with primers specific for the GFP gene, and for two non-targeted regions: Ppap2c and Prkcd, used as negative controls. This figure shows the recovery, expressed as a % of input.</small></p>
<p class="text-right"><a href="../p/crispr-cas9-polyclonal-antibody" class="details tiny button">Learn more</a></p>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>CRISPR/Cas9 monoclonal antibody 4G10</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/landing-pages/cas9_4g10_fig1.png" width="170" height="302" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Antibody raised against N-terminus of Cas9 nuclease</li>
<li>Validated for western blot, IP and immunofluorescence</li>
</ul>
<p><small><strong>Immunofluorescence</strong>: Hela cells were transiently transfected with a Cas9 expression vector. The cells were fixed in 3.7% formaldehyde, permeabilized in 0.5% Triton-X-100 and blocked in PBS containing 2% BSA. The cells were stained with the Cas9 antibody at 4°C o/n, followed by incubation with an anti mouse secondary antibody coupled to AF488 for 1 h at RT. Nuclei were counter-stained with Hoechst 33342 (right).</small></p>
<p class="text-right"><a href="../p/crispr-cas9-monoclonal-antibody-4g10-50-ug" class="details tiny button">Learn more</a></p>
</div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>CRISPR/Cas9 C-terminal monoclonal antibody</h3>
</div>
<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IP.png" /></div>
<div class="small-8 medium-8 large-8 columns">
<ul>
<li>Antibody raised against C-terminus of Cas9 nuclease</li>
<li>Validated for western blot, IP and immunofluorescence</li>
</ul>
<p><small><strong> Western blot</strong> was performed on 20 μg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9. The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
<p class="text-right"><a href="../p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug" class="details tiny button">Learn more</a></p>
</div>
<div class="small-12 medium-12 large-12 columns">
<h3>Which CRISPR/Cas9 antibody is the best for your application?</h3>
<a name="table"></a>
<table>
<thead>
<tr>
<th>Antibody</th>
<th>WB</th>
<th>IF</th>
<th>IP</th>
<th>ChIP</th>
<th>Antibody raised against</th>
</tr>
</thead>
<tbody>
<tr>
<td><a href="../p/crispr-cas9-monoclonal-antibody-50-ug-25-μl"><strong class="diacol">CRISPR/Cas9 monoclonal antibody</strong></a></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><span class="diacol">N-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-polyclonal-antibody">CRISPR/Cas9 polyclonal antibody</a></td>
<td>++</td>
<td>++</td>
<td>++</td>
<td><strong class="diacol">+++</strong></td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-monoclonal-antibody-4g10-50-ug">CRISPR/Cas9 monoclonal antibody 4G10</a></td>
<td>+++</td>
<td>+++</td>
<td>++</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="../p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug-23-ul"><strong class="diacol">CRISPR/Cas9 C-terminal monoclonal antibody</strong></a> <span class="label alert" style="font-size: 0.9rem;">NEW!</span></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">+++</strong></td>
<td><strong class="diacol">no</strong></td>
<td><strong class="diacol">+</strong></td>
<td><span class="diacol">C-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-c-terminal-monoclonal-antibody-50-ug">CRISPR/Cas9 C-terminal monoclonal antibody</a></td>
<td>++</td>
<td>++</td>
<td>+</td>
<td>no</td>
<td><span class="diacol">C-terminus of Cas9 nuclease</span></td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-monoclonal-antibody-7A9-50-mg">CRISPR/Cas9 monoclonal antibody 7A9</a></td>
<td>++</td>
<td>++</td>
<td>++</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
<tr>
<td><a href="/p/crispr-cas9-hrp-monoclonal-antibody-50-ul">CRISPR/Cas9 - HRP monoclonal antibody 7A9</a></td>
<td>+++</td>
<td>no</td>
<td>no</td>
<td>no</td>
<td>N-terminus of Cas9 nuclease</td>
</tr>
</tbody>
</table>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-WB.png" alt="Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 µg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 & micro;g) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 µg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 µg) is shown in lane 1 and 2. </small></p>
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<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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<p><strong><input name="data[Cart][quantity]" placeholder="1" value="1" min="1" style="width:60px;display:inline" type="number" id="CartQuantity" required="required"/></strong>CRISPR/Cas9 Antibody個カートに追加。</p>
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<h6 style="height:60px">CRISPR/Cas9 polyclonal antibody</h6>
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<div class="small-6 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-fig1.png" /></center></div>
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<p><strong>Figure 1. ChIP using the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing nuclease dead Cas9 and sgRNA targeting a sequence in intron 8 of the GAPDH gene, using the iDeal ChIP-seq kit for transcription factors. A titration consisting of 1, 2, 5 and 10 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) was tested. IgG (2 µg/IP) was used as negative IP control. qPCR was performed with primers specific for the targeted sequence in the GAPDH gene, and for the MYOD1 gene, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
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<div class="small-12 columns"><center>A.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-a.jpg" width="700" /></center><center>B.<img src="https://www.diagenode.com/img/product/antibodies/C15310258-chipseq-b.jpg" width="700" /></center></div>
</div>
<div class="row">
<div class="small-12 columns">
<p><strong>Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against Cas9</strong><br />ChIP was performed on sheared chromatin from 4 million HEK293T cells stably expressing dCas9 and a GAPDH sgRNA cells using 2 µl of the Diagenode antibody against Cas9 (cat. No. C15310258) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. 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 ChIP-seq profile in a region of chromosome 12 surrounding the GAPDH gene (fig 2B) and in a region of chromosome 2 surrounding an off-target peak in the YIPF4 gene.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-WB.png" /></center></div>
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<p><strong>Figure 3. Western blot analysis using the Diagenode antibody directed against Cas9</strong><br />Western blot was performed on protein extracts from HEK293 cells transfected with dCas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15310258). The antibody was diluted 1:5,000. The marker is shown on the left, the position of the Cas9 protein is indicated on the right.</p>
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<div class="small-4 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-IP.png" /></center></div>
<div class="small-8 columns">
<p><strong>Figure 4. IP using the Diagenode antibody directed against Cas9</strong><br />IP was performed on whole cell extracts (500 µg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 1 µl of the Diagenode antibody against Cas9 (cat. No. C15310258). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (25 µg) is shown in lane 1 and 2.</p>
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<div class="small-12 columns"><center><img src="https://www.diagenode.com/img/product/antibodies/C15310258-CRISPR-IF.png" width="500" /></center></div>
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<div class="small-12 columns">
<p><strong>Figure 5. Immunofluorescence using the Diagenode antibody directed against Cas9</strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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 antibody (cat. No. C15310258) diluted 1:1000, followed by incubation with a goat anti-rabbit secondary antibody coupled to AF594. Nuclei were counter-stained with Hoechst 33342. Figure 5 shows the result in the presence (left) or absence (right) of doxycycline.</p>
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'info2' => '<p style="text-align: justify;">CRISPR systems are adaptable immune mechanisms which are present in many bacteria to protect themselves from foreign nucleic acids, such as viruses, transposable elements or plasmids. Recently, the <strong>CRISPR/Cas9</strong> (CRISPR-associated protein 9 nuclease, UniProtKB/Swiss-Prot entry Q99ZW2) system from S. pyogenes has been adapted for inducing sequence-specific double stranded breaks and targeted genome editing. This system is unique and flexible due to its dependence on RNA as the moiety that targets the nuclease to a desired DNA sequence and can be used induce indel mutations, specific sequence replacements or insertions and large deletions or genomic rearrangements at any desired location in the genome. In addition, Cas9 can also be used to mediate upregulation of specific endogenous genes or to alter histone modifications or DNA methylation.</p>',
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<p><small><strong> Figure 1. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on protein extracts from HeLa cells transfected with Cas9 using the Diagenode antibody against CRISPR/Cas9 (cat. No. C15200223). The antibody was used at different dilutions. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 2. Western blot analysis using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />Western blot was performed on 20 μg protein extracts from Cas9 expressing HeLa cells (lane 1) and on negative control HeLa cells (lane 2) with the Diagenode antibody against Cas9 (cat. No. C15200223). The antibody was diluted 1:4,000. The marker is shown on the left, position of the Cas9 protein is indicated on the right. </small></p>
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<p><small><strong> Figure 3. IP using the Diagenode monoclonal antibody directed against Cas9 </strong><br />IP was performed on whole cell extracts (250 μg) from HeLa cells transfected with a Cas9 expression vector (lane 1 and 3), or untransfected cells (lane 2 and 4) using 13 μg of the Diagenode antibody against Cas9 (cat. No. C15200223). The immunoprecipitated proteins were subsequently analysed by Western blot. Lane 3 and 4 show the result of the IP, the input (15 μg) is shown in lane 1 and 2. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200223-IF.png" alt="IP" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong> Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9 </strong><br />HeLa cells expressing Cas9 under the control of the tight TRE promoter 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. C15200223) diluted 1:200, followed by incubation with a donkey anti-mouse secondary antibody coupled to AF488. Nuclei were counter-stained with Hoechst 33342. Figure 4 shows the result in the presence (left) or absence (right) of doxycycline. </small></p>
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