5-methylcytosine (5-mC) Antibody for ICC/IF

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	'meta_description' => ' 5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF  validated  in MeDIP, DB, IF, and FISH. Batch-specific data available on the website. ',
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			'name' => ' 5-methylcytosine (5-mC) Antibody for ICC/IF ',
			'description' => '<p><span>Monoclonal antibody raised in mouse against 5-mC (5-methylcytosine) conjugated to BSA.&nbsp;</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_DotBlot.png" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </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/C15200003-IF-1.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<div class="row">
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<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
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<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<div class="small-5 columns">
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<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<td>MeDIP <sup>*</sup></td>
<td>1-2 &micro;g per IP</td>
<td></td>
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<td>Dot Blotting **</td>
<td>1:600</td>
<td>Fig 1</td>
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<td>Immunofluorescence</td>
<td>1:1,000</td>
<td>Fig 2, 3</td>
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<td>FISH</td>
<td>1:200 - 1:1,000</td>
<td>Fig 4, 5</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 &micro;g per IP.</small></p>
<p><small><sup>**</sup> Dot blot was only performed to demonstrate the specificity. This antibody is not recommended for dot blot on biological samples</small></p>',
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		'name' => ' 5-methylcytosine (5-mC) Antibody for ICC/IF ',
		'description' => '<p><span>Monoclonal antibody raised in mouse against 5-mC (5-methylcytosine) conjugated to BSA.&nbsp;</span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_DotBlot.png" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-1.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
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<div class="row">
<div class="small-4 columns">
<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
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<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown 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/C15200003_FISH.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-7 columns">
<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
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		'lot' => '003',
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<th>Suggested dilution</th>
<th>References</th>
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<tr>
<td>MeDIP <sup>*</sup></td>
<td>1-2 &micro;g per IP</td>
<td></td>
</tr>
<tr>
<td>Dot Blotting **</td>
<td>1:600</td>
<td>Fig 1</td>
</tr>
<tr>
<td>Immunofluorescence</td>
<td>1:1,000</td>
<td>Fig 2, 3</td>
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<tr>
<td>FISH</td>
<td>1:200 - 1:1,000</td>
<td>Fig 4, 5</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 &micro;g per IP.</small></p>
<p><small><sup>**</sup> Dot blot was only performed to demonstrate the specificity. This antibody is not recommended for dot blot on biological samples</small></p>',
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			'name' => ' 5-methylcytosine (5-mC) Antibody for ICC/IF ',
			'description' => '<p><span>Monoclonal antibody raised in mouse against 5-mC (5-methylcytosine) conjugated to BSA.&nbsp;</span></p>',
			'label1' => 'Validation Data',
			'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_DotBlot.png" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-1.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH_2.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
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			'format' => '50 μg/39 μl',
			'catalog_number' => 'C15200003',
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			'sf_code' => 'C15200003-D001-000581',
			'type' => 'FRE',
			'search_order' => '03-Antibody',
			'price_EUR' => '380',
			'price_USD' => '380',
			'price_GBP' => '340',
			'price_JPY' => '59525',
			'price_CNY' => '',
			'price_AUD' => '950',
			'country' => 'ALL',
			'except_countries' => 'None',
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			'last_datasheet_update' => 'July 30, 2020',
			'slug' => '5-mc-monoclonal-antibody-for-icc-if-classic-50-mg-21-ml',
			'meta_title' => '5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF | Diagenode',
			'meta_keywords' => '5-methylcytosine (5-mC),monoclonal antibody ',
			'meta_description' => ' 5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF  validated  in MeDIP, DB, IF, and FISH. Batch-specific data available on the website. ',
			'modified' => '2022-01-05 15:26:15',
			'created' => '2015-06-29 14:08:20'
		),
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			'antibody_id' => null,
			'name' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'description' => '<p><a href="https://www.diagenode.com/files/products/kits/hMeDIP_kit_manual.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p><span>The hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented&nbsp;genomic DNA<span><span>&nbsp;</span>samples for use in genome-wide methylation analysis. It features</span></span><span>&nbsp;a highly specific monoclonal antibody against&nbsp;</span>5-hydroxymethylcytosine (5-hmC) for the immunoprecipitation of hydroxymethylated DNA<span>. It includes&nbsp;control DNA and primers to assess the effiency of the assay.&nbsp;</span>Performing hydroxymethylation profiling with the hMeDIP kit is fast,&nbsp;reliable and&nbsp;highly specific.</p>
<p><em>Looking for hMeDIP-seq protocol? <a href="https://go.diagenode.com/l/928883/2022-01-07/2m1ht" target="_blank" title="Contact us">Contact us</a></em></p>
<p><span></span></p>
<p><span></span></p>',
			'label1' => 'Characteristics',
			'info1' => '<ul style="list-style-type: disc;">
<li><span>Robust enrichment &amp; immunoprecipitation of hydroxymethylated DNA</span></li>
<li>Highly specific monoclonal antibody against 5-hmC<span>&nbsp;for reliable, reproducible results</span></li>
<li>Including&nbsp;control DNA and primers to&nbsp;<span>monitor the&nbsp;efficiency of the assay</span>
<ul style="list-style-type: circle;">
<li>hmeDNA and unmethylated DNA sequences and primer pairs</li>
<li>Mouse primer pairs against Sfi1 targeting hydroxymethylated gene in mouse</li>
</ul>
</li>
<li>Improved single-tube, magnetic bead-based protocol</li>
</ul>',
			'label2' => '',
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			'format' => '16 rxns',
			'catalog_number' => 'C02010031',
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			'sf_code' => 'C02010031-',
			'type' => 'RFR',
			'search_order' => '04-undefined',
			'price_EUR' => '630',
			'price_USD' => '690',
			'price_GBP' => '580',
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			'price_AUD' => '1725',
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			'slug' => 'hmedip-kit-x16-monoclonal-mouse-antibody-16-rxns',
			'meta_title' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'meta_keywords' => '',
			'meta_description' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'modified' => '2023-04-20 16:12:48',
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			'antibody_id' => null,
			'name' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'description' => '<p><span>The Auro hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented&nbsp;genomic DNA&nbsp;samples for use in genome-wide methylation analysis. It features</span><span>&nbsp;a highly specific monoclonal antibody against&nbsp;</span><span>5-hydroxymethylcytosine (5-hmC) for the immunoprecipitation of hydroxymethylated DNA</span><span>. It includes&nbsp;control DNA and primers to assess the effiency of the assay.&nbsp;</span><span>Performing hydroxymethylation profiling with the hMeDIP kit is fast,&nbsp;reliable and&nbsp;highly specific.</span></p>',
			'label1' => ' Characteristics',
			'info1' => '<ul style="list-style-type: disc;">
<li><span>Robust enrichment &amp; immunoprecipitation of hydroxymethylated DNA</span></li>
<li>Highly specific monoclonal antibody against 5-hmC<span>&nbsp;for reliable, reproducible results</span></li>
<li>Including&nbsp;control DNA and primers to&nbsp;<span>monitor the&nbsp;efficiency of the assay</span>
<ul style="list-style-type: circle;">
<li>5-hmC, 5-mC and unmethylated DNA sequences and primer pairs</li>
<li>Mouse primer pairs against Sfi1 targeting hydroxymethylated gene in mouse</li>
</ul>
</li>
</ul>
<ul style="list-style-type: disc;">
<li>Improved single-tube, magnetic bead-based protocol</li>
</ul>',
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			'format' => '16 rxns',
			'catalog_number' => 'C02010034',
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			'sf_code' => 'C02010034-',
			'type' => 'RFR',
			'search_order' => '04-undefined',
			'price_EUR' => '630',
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			'price_CNY' => '',
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			'slug' => 'auto-hmedip-kit-x16-monoclonal-mouse-antibody-16-rxns',
			'meta_title' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'meta_keywords' => '',
			'meta_description' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'modified' => '2021-01-18 10:37:19',
			'created' => '2015-06-29 14:08:20',
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			'id' => '1880',
			'antibody_id' => null,
			'name' => 'MagMeDIP Kit',
			'description' => '<p><a href="https://www.diagenode.com/files/products/kits/magmedip-kit-manual-C02010020-21.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>Perform <strong>MeDIP</strong> (<strong>Me</strong>thylated <strong>D</strong>NA <strong>I</strong>mmuno<strong>p</strong>recipitation) followed by qPCR or NGS to estimate DNA methylation status of your sample using a highly sensitive 5-methylcytosine antibody. Our MagMeDIP kit contains high quality reagents to get the highest enrichment of methylated DNA with an optimized user-friendly protocol.</p>
<h3><span>Features</span></h3>
<ul>
<li>Starting DNA amount: <strong>10 ng &ndash; 1 &micro;g</strong></li>
<li>Content: <strong>all reagents included</strong> for DNA extraction, immunoprecipitation (including the 5-mC antibody, spike-in controls and their corresponding qPCR primer pairs) as well as DNA isolation after IP.</li>
<li>Application: <strong>qPCR</strong> and <strong>NGS</strong></li>
<li>Robust method, <strong>superior enrichment</strong>, and easy-to-use protocol</li>
<li><strong>High reproducibility</strong> between replicates and repetitive experiments</li>
<li>Compatible with <strong>all species&nbsp;</strong></li>
</ul>
<p>&nbsp;</p>
<div class="small-12 medium-4 large-4 columns"><center></center><center></center><center></center><center><a href="https://www.ncbi.nlm.nih.gov/pubmed/30429608" target="_blank"><img src="https://www.diagenode.com/img/banners/banner-nature-publication-580.png" alt="Click here to read more about MeDIP " caption="false" width="80%" /></a></center></div>
<div class="small-12 medium-8 large-8 columns">
<h3 style="text-align: justify;">Sensitive tumour detection and classification using plasma cell-free DNA methylomes<br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/30429608" target="_blank">Read the publication</a></h3>
<h3 class="c-article-title u-h1" data-test="article-title" itemprop="name headline" style="text-align: justify;">Preparation of cfMeDIP-seq libraries for methylome profiling of plasma cell-free DNA<br /><a href="https://www.nature.com/articles/s41596-019-0202-2" target="_blank" title="cfMeDIP-seq Nature Method">Read the method</a></h3>
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<p></p>
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<h3></h3>',
			'label1' => 'MagMeDIP workflow',
			'info1' => '<p>DNA methylation occurs primarily as 5-methylcytosine (5-mC), and the Diagenode MagMeDIP Kit takes advantage of a specific antibody targeting this 5-mC to immunoprecipitate methylated DNA, which can be thereafter directly analyzed by qPCR or Next-Generation Sequencing (NGS).</p>
<h3><span>How it works</span></h3>
<p>In brief, after the cell collection and lysis, the genomic DNA is extracted, sheared, and then denatured. In the next step the antibody directed against 5 methylcytosine and antibody binding beads are used for immunoselection and immunoprecipitation of methylated DNA fragments. Then, the IP&rsquo;d methylated DNA is isolated and can be used for any subsequent analysis as qPCR, amplification, hybridization on microarrays or next generation sequencing.</p>
<center><img src="https://www.diagenode.com/img/product/kits/MagMeDIP-workflow.png" width="70%" alt="5-methylcytosine" caption="false" /></center>
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			'label2' => 'MeDIP-qPCR',
			'info2' => '<p>The kit MagMeDIP contains all reagents necessary for a complete MeDIP-qPCR workflow. Two MagMeDIP protocols have been validated: for manual processing as well as for automated processing, using the Diagenode&rsquo;s IP-Star Compact Automated System (please refer to the kit manual).</p>
<ul>
<li><strong>Complete kit</strong> including DNA extraction module, IP antibody and reagents, DNA isolation buffer</li>
<li><strong>Quality control of the IP:</strong> due to methylated and unmethylated DNA spike-in controls and their associated qPCR primers</li>
<li><strong>Easy to use</strong> with user-friendly magnetic beads and rack</li>
<li><strong>Highly validated protocol</strong></li>
<li>Automated protocol supplied</li>
</ul>
<center><img src="https://www.diagenode.com/img/product/kits/fig1-magmedipkit.png" width="85%" alt="Methylated DNA Immunoprecipitation" caption="false" /></center>
<p style="font-size: 0.9em;"><em><strong>Figure 1.</strong> Immunoprecipitation results obtained with Diagenode MagMeDIP Kit</em></p>
<p style="font-size: 0.9em;">MeDIP assays were performed manually using 1 &micro;g or 50 ng gDNA from blood cells with the MagMeDIP kit (Diagenode). The IP was performed with the Methylated and Unmethylated spike-in controls included in the kit, together with the human DNA samples. The DNA was isolated/purified using DIB. Afterwards, qPCR was performed using the primer pairs included in this kit.</p>
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			'label3' => 'MeDIP-seq',
			'info3' => '<p>For DNA methylation analysis on the whole genome, MagMeDIP kit can be coupled with Next-Generation Sequencing. To perform MeDIP-sequencing we recommend the following strategy:</p>
<ul style="list-style-type: circle;">
<li>Choose a library preparation solution which is compatible with the starting amount of DNA you are planning to use (from 10 ng to 1 &mu;g). It can be a home-made solution or a commercial one.</li>
<li>Choose the indexing system that fits your needs considering the following features:</li>
<ul>
<ul>
<ul>
<li>Single-indexing, combinatorial dual-indexing or unique dual-indexing</li>
<li>Number of barcodes</li>
<li>Full-length adaptors containing the barcodes or barcoding at the final amplification step</li>
<li>Presence / absence of Unique Molecular Identifiers (for PCR duplicates removal)</li>
</ul>
</ul>
</ul>
<li>Standard library preparation protocols are compatible with double-stranded DNA only, therefore the first steps of the library preparation (end repair, A-tailing, adaptor ligation and clean-up) will have to be performed on sheared DNA, before the IP.</li>
</ul>
<p style="padding-left: 30px;"><strong>CAUTION:</strong> As the immunoprecipitation step occurs at the middle of the library preparation workflow, single-tube solutions for library preparation are usually not compatible with MeDIP-sequencing.</p>
<ul style="list-style-type: circle;">
<li>For DNA isolation after the IP, we recommend using the <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24" title="IPure kit v2">IPure kit v2</a> (available separately, Cat. No. C03010014) instead of DNA isolation Buffer.</li>
</ul>
<ul style="list-style-type: circle;">
<li>Perform library amplification after the DNA isolation following the standard protocol of the chosen library preparation solution.</li>
</ul>
<h3><span>MeDIP-seq workflow</span></h3>
<center><img src="https://www.diagenode.com/img/product/kits/MeDIP-seq-workflow.png" width="110%" alt="MagMeDIP qPCR Kit x10 workflow" caption="false" /></center>
<h3><span>Example of results</span></h3>
<center><img src="https://www.diagenode.com/img/product/kits/medip-specificity.png" alt="MagMeDIP qPCR Kit Result" caption="false" width="951" height="488" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 1. qPCR analysis of external spike-in DNA controls (methylated and unmethylated) after IP.</strong> Samples were prepared using 1&mu;g &ndash; 100ng -10ng sheared human gDNA with the MagMeDIP kit (Diagenode) and a commercially available library prep kit. DNA isolation after IP has been performed with IPure kit V2 (Diagenode).</p>
<p></p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/medip-saturation-analysis.png" alt=" MagMeDIP kit " caption="false" width="951" height="461" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 2. Saturation analysis.</strong> Clean reads were aligned to the human genome (hg19) using Burrows-Wheeler aligner (BWA) algorithm after which duplicated and unmapped reads were removed resulting in a mapping efficiency &gt;98% for all samples. Quality and validity check of the mapped MeDIP-seq data was performed using MEDIPS R package. Saturation plots show that all sets of reads have sufficient complexity and depth to saturate the coverage profile of the reference genome and that this is reproducible between replicates and repetitive experiments (data shown for 50 ng gDNA input: left panel = replicate a, right panel = replicate b).</p>
<p></p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/medip-libraries-prep.png" alt="MagMeDIP x10 " caption="false" width="951" height="708" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 3. Sequencing profiles of MeDIP-seq libraries prepared from different starting amounts of sheared gDNA on the positive and negative methylated control regions.</strong> MeDIP-seq libraries were prepared from decreasing starting amounts of gDNA (1 &mu;g (green), 50 ng (red), and 10ng (blue)) originating from human blood with the MagMeDIP kit (Diagenode) and a commercially available library prep kit. DNA isolation after IP has been performed with IPure kit V2 (Diagenode). IP and corresponding INPUT samples were sequenced on Illumina NovaSeq SP with 2x50 PE reads. The reads were mapped to the human genome (hg19) with bwa and the alignments were loaded into IGV (the tracks use an identical scale). The top IGV figure shows the TSH2B (also known as H2BC1) gene (marked by blue boxes in the bottom track) and its surroundings. The TSH2B gene is coding for a histone variant that does not occur in blood cells, and it is known to be silenced by methylation. Accordingly, we see a high coverage in the vicinity of this gene. The bottom IGV figure shows the GADPH locus (marked by blue boxes in the bottom track) and its surroundings. The GADPH gene is a highly active transcription region and should not be methylated, resulting in no reads accumulation following MeDIP-seq experiment.</p>
<p></p>
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			'description' => '<p>The MethylCap kit allows to specifically capture DNA fragments containing methylated CpGs. The assay is based on the affinity purification of methylated DNA using methyl-CpG-binding domain (MBD) of human MeCP2 protein. The procedure has been adapted to both manual process or <a href="https://www.diagenode.com/en/p/sx-8g-ip-star-compact-automated-system-1-unit">IP-Star&reg; Compact Automated System</a>. Libraries of captured methylated DNA can be prepared for next-generation sequencing (NGS) by combining MBD technology with the <a href="https://www.diagenode.com/en/p/microplex-lib-prep-kit-v3-48-rxns">MicroPlex Library Preparation Kit v3</a>.</p>',
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<li><strong>Differential fractionation</strong> of methylated DNA by CpG density (3 eluted fractions)</li>
<li><strong>On-day protocol</strong></li>
<li><strong>NGS compatibility</strong></li>
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<h3>MBD-seq allows for detection of genomic regions with different CpG density</h3>
<p><img src="https://www.diagenode.com/img/product/kits/mbd_results1.png" alt="MBD-sequencing results have been validated by bisulfite sequencing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong></strong></p>
<p><strong></strong><strong>F</strong><strong>igure 1.</strong> Using the MBD approach, two methylated regions were detected in different elution fractions according to their methylated CpG density (A). Low, Medium and High refer to the sequenced DNA from different elution fractions with increasing salt concentration. Methylated patterns of these two different methylated regions were validated by bisulfite conversion assay (B).</p>',
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			'description' => '<p>The Auto MethylCap kit allows to specifically capture DNA fragments containing methylated CpGs. The assay is based on the affinity purification of methylated DNA using methyl-CpG-binding domain (MBD) of human MeCP2 protein. This procedure has been optimized to perform automated immunoprecipitation of chromatin using the <a href="https://www.diagenode.com/en/p/sx-8g-ip-star-compact-automated-system-1-unit">IP-Star&reg; Compact Automated System</a> enabling highly reproducible results and allowing for high throughput.</p>',
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<li><strong>Automation compatibility</strong><strong></strong>
<h3>MBD-seq allows for detection of genomic regions with different CpG density</h3>
<p><img src="https://www.diagenode.com/img/product/kits/mbd_results1.png" alt="MBD-sequencing results have been validated by bisulfite sequencing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>F</strong><strong>igure 1.</strong><span>&nbsp;</span>Using the MBD approach, two methylated regions were detected in different elution fractions according to their methylated CpG density (A). Low, Medium and High refer to the sequenced DNA from different elution fractions with increasing salt concentration. Methylated patterns of these two different methylated regions were validated by bisulfite conversion assay (B).<br /><strong></strong></p>
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<p style="text-align: center;"><strong>Make your Bisulfite conversion now in only 60 minutes !</strong></p>
<p>Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion reagent is added directly to DNA, requires no intermediate steps, and results in high yields of DNA ready for downstream analysis methods including PCR and Next-Generation Sequencing.</p>',
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			'description' => '<p><strong>Immunofluorescence</strong>:</p>
<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&deg;C, permeabilized with acetone at -20&deg;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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			'meta_description' => '5-Methylcytosine(5-mC) monoclonal antibody for fish ',
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			'name' => 'DNA modifications',
			'description' => '<p><span style="font-weight: 400;">T</span><span style="font-weight: 400;">he pattern of <strong>DNA modifications</strong> is critical for genome stability and the control of gene expression in the cell. Methylation of 5-cytosine (5-mC), one of the best-studied epigenetic marks, is carried out by the <strong>DNA methyltransferases</strong> DNMT3A and B and DNMT1. DNMT3A and DNMT3B are responsible for </span><i><span style="font-weight: 400;">de novo</span></i><span style="font-weight: 400;"> DNA methylation, whereas DNMT1 maintains existing methylation. 5-mC undergoes active demethylation which is performed by the <strong>Ten-Eleven Translocation</strong> (TET) familly of DNA hydroxylases. The latter consists of 3 members TET1, 2 and 3. All 3 members catalyze the conversion of <strong>5-methylcytosine</strong> (5-mC) into <strong>5-hydroxymethylcytosine</strong> (5-hmC), and further into <strong>5-formylcytosine</strong> (5-fC) and <strong>5-carboxycytosine</strong> (5-caC). 5-fC and 5-caC can be converted to unmodified cytosine by <strong>Thymine DNA Glycosylase</strong> (TDG). It is not yet clear if 5-hmC, 5-fC and 5-caC have specific functions or are simply intermediates in the demethylation of 5-mC.</span></p>
<p><span style="font-weight: 400;">DNA methylation is generally considered as a repressive mark and is usually associated with gene silencing. It is essential that the balance between DNA methylation and demethylation is precisely maintained. Dysregulation of DNA methylation may lead to many different human diseases and is often observed in cancer cells.</span></p>
<p><span style="font-weight: 400;">Diagenode offers highly validated antibodies against different proteins involved in DNA modifications as well as against the modified bases allowing the study of all steps and intermediates in the DNA methylation/demethylation pathway:</span></p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/dna-methylation.jpg" height="599" width="816" /></p>
<p><strong>Diagenode exclusively sources the original 5-methylcytosine monoclonal antibody (clone 33D3).</strong></p>
<p>Check out the list below to see all proposed antibodies for DNA modifications.</p>
<p>Diagenode&rsquo;s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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			'meta_description' => 'Diagenode offers Monoclonal and Polyclonal antibodies for DNA Methylation. The pattern of DNA modifications is critical for genome stability and the control of gene expression in the cell. ',
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			'name' => 'All antibodies',
			'description' => '<p><span style="font-weight: 400;">All Diagenode&rsquo;s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode&rsquo;s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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			'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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			'name' => 'Developmental mRNA mC landscape and regulatory innovations of massivemC modification of maternal mRNAs in animals.',
			'authors' => 'Liu  J. et al.',
			'description' => '<p>mC is one of the longest-known RNA modifications, however, its developmental dynamics, functions, and evolution in mRNAs remain largely unknown. Here, we generate quantitative mRNA mC maps at different stages of development in 6 vertebrate and invertebrate species and find convergent and unexpected massive methylation of maternal mRNAs mediated by NSUN2 and NSUN6. Using Drosophila as a model, we reveal that embryos lacking maternal mRNA mC undergo cell cycle delays and fail to timely initiate maternal-to-zygotic transition, implying the functional importance of maternal mRNA mC. From invertebrates to the lineage leading to humans, two waves of mC regulatory innovations are observed: higher animals gain cis-directed NSUN2-mediated mC sites at the 5' end of the mRNAs, accompanied by the emergence of more structured 5'UTR regions; humans gain thousands of trans-directed NSUN6-mediated mC sites enriched in genes regulating the mitotic cell cycle. Collectively, our studies highlight the existence and regulatory innovations of a mechanism of early embryonic development and provide key resources for elucidating the role of mRNA mC in biology and disease.</p>',
			'date' => '2022-05-01',
			'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/35513466',
			'doi' => '10.1038/s41467-022-30210-0',
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			'name' => 'Detection of genetic variation and base modifications at base-pairresolution on both DNA and RNA.',
			'authors' => 'Wang, Zhen et al.',
			'description' => '<p>Accurate decoding of nucleic acid variation is critical to understand the complexity and regulation of genome function. Here we use a single-molecule magnetic tweezer (MT) platform to identify sequence variation and map a range of important epigenetic base modifications with high sensitivity, specificity, and precision in the same single molecules of DNA or RNA. We have also developed a highly specific amplification-free CRISPR-Cas enrichment strategy to isolate genomic regions from native DNA. We demonstrate enrichment of DNA from both E. coli and the FMR1 5'UTR coming from cells derived from a Fragile X carrier. From these kilobase-length enriched molecules we could characterize the differential levels of adenine and cytosine base modifications on E. coli, and the repeat expansion length and methylation status of FMR1. Together these results demonstrate that our platform can detect a variety of genetic, epigenetic, and base modification changes concomitantly within the same single molecules.</p>',
			'date' => '2021-01-01',
			'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/33514840',
			'doi' => '10.1038/s42003-021-01648-7',
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			'name' => 'Transcriptome-wide distribution and function of RNA hydroxymethylcytosine',
			'authors' => 'Delatte B, Wang F, Ngoc LV, Collignon E, Bonvin E, Deplus R, Calonne E, Hassabi B, Putmans P, Awe S, Wetzel C, Kreher J, Soin R, Creppe C, Limbach PA, Gueydan C, Kruys V, Brehm A, Minakhina S, Defrance M, Steward R, Fuks F.',
			'description' => '<p>Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.</p>',
			'date' => '2016-01-15',
			'pmid' => 'http://pubmed.gov/26816380',
			'doi' => '10.1126/science.aac5253',
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	'authors' => 'Delatte B, Wang F, Ngoc LV, Collignon E, Bonvin E, Deplus R, Calonne E, Hassabi B, Putmans P, Awe S, Wetzel C, Kreher J, Soin R, Creppe C, Limbach PA, Gueydan C, Kruys V, Brehm A, Minakhina S, Defrance M, Steward R, Fuks F',
	'description' => '<p>Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.</p>',
	'date' => '2016-01-15',
	'pmid' => 'http://www.pubmed.org/26816380',
	'doi' => '10.1126/science.aac5253',
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include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118

Notice (8): Undefined variable: header [APP/View/Products/view.ctp, line 755]
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	'meta_keywords' => '5-methylcytosine (5-mC),monoclonal antibody ',
	'meta_description' => ' 5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF  validated  in MeDIP, DB, IF, and FISH. Batch-specific data available on the website. ',
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			'name' => ' 5-methylcytosine (5-mC) Antibody for ICC/IF ',
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<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-1.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-8 columns">
<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
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<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>MeDIP <sup>*</sup></td>
<td>1-2 &micro;g per IP</td>
<td></td>
</tr>
<tr>
<td>Dot Blotting **</td>
<td>1:600</td>
<td>Fig 1</td>
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<tr>
<td>Immunofluorescence</td>
<td>1:1,000</td>
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<tr>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 &micro;g per IP.</small></p>
<p><small><sup>**</sup> Dot blot was only performed to demonstrate the specificity. This antibody is not recommended for dot blot on biological samples</small></p>',
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<div class="small-7 columns">
<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH_2.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
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		'label2' => '',
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		'format' => '50 μg/39 μl',
		'catalog_number' => 'C15200003',
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		'sf_code' => 'C15200003-D001-000581',
		'type' => 'FRE',
		'search_order' => '03-Antibody',
		'price_EUR' => '380',
		'price_USD' => '380',
		'price_GBP' => '340',
		'price_JPY' => '59525',
		'price_CNY' => '',
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		'slug' => '5-mc-monoclonal-antibody-for-icc-if-classic-50-mg-21-ml',
		'meta_title' => '5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF | Diagenode',
		'meta_keywords' => '5-methylcytosine (5-mC),monoclonal antibody ',
		'meta_description' => ' 5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF  validated  in MeDIP, DB, IF, and FISH. Batch-specific data available on the website. ',
		'modified' => '2022-01-05 15:26:15',
		'created' => '2015-06-29 14:08:20',
		'locale' => 'eng'
	),
	'Antibody' => array(
		'host' => '*****',
		'id' => '30',
		'name' => '5-mC monoclonal antibody for ICC/IF',
		'description' => 'Monoclonal antibody raised in mouse against 5-mC (5-methyl Cytidine) conjugated to BSA. ',
		'clonality' => '',
		'isotype' => 'IgG1',
		'lot' => '003',
		'concentration' => '1.3 &micro;g/&micro;l',
		'reactivity' => 'Human, mouse, drosophila, other (wide range): positive.',
		'type' => 'Monoclonal',
		'purity' => 'Protein A purified monoclonal antibody.',
		'classification' => 'Classic',
		'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>MeDIP <sup>*</sup></td>
<td>1-2 &micro;g per IP</td>
<td></td>
</tr>
<tr>
<td>Dot Blotting **</td>
<td>1:600</td>
<td>Fig 1</td>
</tr>
<tr>
<td>Immunofluorescence</td>
<td>1:1,000</td>
<td>Fig 2, 3</td>
</tr>
<tr>
<td>FISH</td>
<td>1:200 - 1:1,000</td>
<td>Fig 4, 5</td>
</tr>
</tbody>
</table>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 &micro;g per IP.</small></p>
<p><small><sup>**</sup> Dot blot was only performed to demonstrate the specificity. This antibody is not recommended for dot blot on biological samples</small></p>',
		'storage_conditions' => 'Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.',
		'storage_buffer' => 'PBS containing 0.05% azide.',
		'precautions' => 'This product is for research use only. Not for use in diagnostic or therapeutic procedures.',
		'uniprot_acc' => '',
		'slug' => '',
		'meta_keywords' => '',
		'meta_description' => '',
		'modified' => '2020-07-30 15:53:25',
		'created' => '0000-00-00 00:00:00',
		'select_label' => '30 - 5-mC monoclonal antibody for ICC/IF (003 - 1.3 &micro;g/&micro;l - Human, mouse, drosophila, other (wide range): positive. - Protein A purified monoclonal antibody. - Mouse)'
	),
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			'created' => '2020-11-18 16:07:10'
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	),
	'Group' => array(
		'Group' => array(
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			'id' => '1961',
			'antibody_id' => '30',
			'name' => ' 5-methylcytosine (5-mC) Antibody for ICC/IF ',
			'description' => '<p><span>Monoclonal antibody raised in mouse against 5-mC (5-methylcytosine) conjugated to BSA.&nbsp;</span></p>',
			'label1' => 'Validation Data',
			'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_DotBlot.png" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-1.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
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<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH_2.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
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			'meta_title' => '5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF | Diagenode',
			'meta_keywords' => '5-methylcytosine (5-mC),monoclonal antibody ',
			'meta_description' => ' 5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF  validated  in MeDIP, DB, IF, and FISH. Batch-specific data available on the website. ',
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<p><span>The hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented&nbsp;genomic DNA<span><span>&nbsp;</span>samples for use in genome-wide methylation analysis. It features</span></span><span>&nbsp;a highly specific monoclonal antibody against&nbsp;</span>5-hydroxymethylcytosine (5-hmC) for the immunoprecipitation of hydroxymethylated DNA<span>. It includes&nbsp;control DNA and primers to assess the effiency of the assay.&nbsp;</span>Performing hydroxymethylation profiling with the hMeDIP kit is fast,&nbsp;reliable and&nbsp;highly specific.</p>
<p><em>Looking for hMeDIP-seq protocol? <a href="https://go.diagenode.com/l/928883/2022-01-07/2m1ht" target="_blank" title="Contact us">Contact us</a></em></p>
<p><span></span></p>
<p><span></span></p>',
			'label1' => 'Characteristics',
			'info1' => '<ul style="list-style-type: disc;">
<li><span>Robust enrichment &amp; immunoprecipitation of hydroxymethylated DNA</span></li>
<li>Highly specific monoclonal antibody against 5-hmC<span>&nbsp;for reliable, reproducible results</span></li>
<li>Including&nbsp;control DNA and primers to&nbsp;<span>monitor the&nbsp;efficiency of the assay</span>
<ul style="list-style-type: circle;">
<li>hmeDNA and unmethylated DNA sequences and primer pairs</li>
<li>Mouse primer pairs against Sfi1 targeting hydroxymethylated gene in mouse</li>
</ul>
</li>
<li>Improved single-tube, magnetic bead-based protocol</li>
</ul>',
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			'slug' => 'hmedip-kit-x16-monoclonal-mouse-antibody-16-rxns',
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		(int) 1 => array(
			'id' => '1885',
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			'name' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'description' => '<p><span>The Auro hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented&nbsp;genomic DNA&nbsp;samples for use in genome-wide methylation analysis. It features</span><span>&nbsp;a highly specific monoclonal antibody against&nbsp;</span><span>5-hydroxymethylcytosine (5-hmC) for the immunoprecipitation of hydroxymethylated DNA</span><span>. It includes&nbsp;control DNA and primers to assess the effiency of the assay.&nbsp;</span><span>Performing hydroxymethylation profiling with the hMeDIP kit is fast,&nbsp;reliable and&nbsp;highly specific.</span></p>',
			'label1' => ' Characteristics',
			'info1' => '<ul style="list-style-type: disc;">
<li><span>Robust enrichment &amp; immunoprecipitation of hydroxymethylated DNA</span></li>
<li>Highly specific monoclonal antibody against 5-hmC<span>&nbsp;for reliable, reproducible results</span></li>
<li>Including&nbsp;control DNA and primers to&nbsp;<span>monitor the&nbsp;efficiency of the assay</span>
<ul style="list-style-type: circle;">
<li>5-hmC, 5-mC and unmethylated DNA sequences and primer pairs</li>
<li>Mouse primer pairs against Sfi1 targeting hydroxymethylated gene in mouse</li>
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<li>Improved single-tube, magnetic bead-based protocol</li>
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			'meta_description' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
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		(int) 2 => array(
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			'name' => 'MagMeDIP Kit',
			'description' => '<p><a href="https://www.diagenode.com/files/products/kits/magmedip-kit-manual-C02010020-21.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>Perform <strong>MeDIP</strong> (<strong>Me</strong>thylated <strong>D</strong>NA <strong>I</strong>mmuno<strong>p</strong>recipitation) followed by qPCR or NGS to estimate DNA methylation status of your sample using a highly sensitive 5-methylcytosine antibody. Our MagMeDIP kit contains high quality reagents to get the highest enrichment of methylated DNA with an optimized user-friendly protocol.</p>
<h3><span>Features</span></h3>
<ul>
<li>Starting DNA amount: <strong>10 ng &ndash; 1 &micro;g</strong></li>
<li>Content: <strong>all reagents included</strong> for DNA extraction, immunoprecipitation (including the 5-mC antibody, spike-in controls and their corresponding qPCR primer pairs) as well as DNA isolation after IP.</li>
<li>Application: <strong>qPCR</strong> and <strong>NGS</strong></li>
<li>Robust method, <strong>superior enrichment</strong>, and easy-to-use protocol</li>
<li><strong>High reproducibility</strong> between replicates and repetitive experiments</li>
<li>Compatible with <strong>all species&nbsp;</strong></li>
</ul>
<p>&nbsp;</p>
<div class="small-12 medium-4 large-4 columns"><center></center><center></center><center></center><center><a href="https://www.ncbi.nlm.nih.gov/pubmed/30429608" target="_blank"><img src="https://www.diagenode.com/img/banners/banner-nature-publication-580.png" alt="Click here to read more about MeDIP " caption="false" width="80%" /></a></center></div>
<div class="small-12 medium-8 large-8 columns">
<h3 style="text-align: justify;">Sensitive tumour detection and classification using plasma cell-free DNA methylomes<br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/30429608" target="_blank">Read the publication</a></h3>
<h3 class="c-article-title u-h1" data-test="article-title" itemprop="name headline" style="text-align: justify;">Preparation of cfMeDIP-seq libraries for methylome profiling of plasma cell-free DNA<br /><a href="https://www.nature.com/articles/s41596-019-0202-2" target="_blank" title="cfMeDIP-seq Nature Method">Read the method</a></h3>
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			'info1' => '<p>DNA methylation occurs primarily as 5-methylcytosine (5-mC), and the Diagenode MagMeDIP Kit takes advantage of a specific antibody targeting this 5-mC to immunoprecipitate methylated DNA, which can be thereafter directly analyzed by qPCR or Next-Generation Sequencing (NGS).</p>
<h3><span>How it works</span></h3>
<p>In brief, after the cell collection and lysis, the genomic DNA is extracted, sheared, and then denatured. In the next step the antibody directed against 5 methylcytosine and antibody binding beads are used for immunoselection and immunoprecipitation of methylated DNA fragments. Then, the IP&rsquo;d methylated DNA is isolated and can be used for any subsequent analysis as qPCR, amplification, hybridization on microarrays or next generation sequencing.</p>
<center><img src="https://www.diagenode.com/img/product/kits/MagMeDIP-workflow.png" width="70%" alt="5-methylcytosine" caption="false" /></center>
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			'info2' => '<p>The kit MagMeDIP contains all reagents necessary for a complete MeDIP-qPCR workflow. Two MagMeDIP protocols have been validated: for manual processing as well as for automated processing, using the Diagenode&rsquo;s IP-Star Compact Automated System (please refer to the kit manual).</p>
<ul>
<li><strong>Complete kit</strong> including DNA extraction module, IP antibody and reagents, DNA isolation buffer</li>
<li><strong>Quality control of the IP:</strong> due to methylated and unmethylated DNA spike-in controls and their associated qPCR primers</li>
<li><strong>Easy to use</strong> with user-friendly magnetic beads and rack</li>
<li><strong>Highly validated protocol</strong></li>
<li>Automated protocol supplied</li>
</ul>
<center><img src="https://www.diagenode.com/img/product/kits/fig1-magmedipkit.png" width="85%" alt="Methylated DNA Immunoprecipitation" caption="false" /></center>
<p style="font-size: 0.9em;"><em><strong>Figure 1.</strong> Immunoprecipitation results obtained with Diagenode MagMeDIP Kit</em></p>
<p style="font-size: 0.9em;">MeDIP assays were performed manually using 1 &micro;g or 50 ng gDNA from blood cells with the MagMeDIP kit (Diagenode). The IP was performed with the Methylated and Unmethylated spike-in controls included in the kit, together with the human DNA samples. The DNA was isolated/purified using DIB. Afterwards, qPCR was performed using the primer pairs included in this kit.</p>
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			'label3' => 'MeDIP-seq',
			'info3' => '<p>For DNA methylation analysis on the whole genome, MagMeDIP kit can be coupled with Next-Generation Sequencing. To perform MeDIP-sequencing we recommend the following strategy:</p>
<ul style="list-style-type: circle;">
<li>Choose a library preparation solution which is compatible with the starting amount of DNA you are planning to use (from 10 ng to 1 &mu;g). It can be a home-made solution or a commercial one.</li>
<li>Choose the indexing system that fits your needs considering the following features:</li>
<ul>
<ul>
<ul>
<li>Single-indexing, combinatorial dual-indexing or unique dual-indexing</li>
<li>Number of barcodes</li>
<li>Full-length adaptors containing the barcodes or barcoding at the final amplification step</li>
<li>Presence / absence of Unique Molecular Identifiers (for PCR duplicates removal)</li>
</ul>
</ul>
</ul>
<li>Standard library preparation protocols are compatible with double-stranded DNA only, therefore the first steps of the library preparation (end repair, A-tailing, adaptor ligation and clean-up) will have to be performed on sheared DNA, before the IP.</li>
</ul>
<p style="padding-left: 30px;"><strong>CAUTION:</strong> As the immunoprecipitation step occurs at the middle of the library preparation workflow, single-tube solutions for library preparation are usually not compatible with MeDIP-sequencing.</p>
<ul style="list-style-type: circle;">
<li>For DNA isolation after the IP, we recommend using the <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24" title="IPure kit v2">IPure kit v2</a> (available separately, Cat. No. C03010014) instead of DNA isolation Buffer.</li>
</ul>
<ul style="list-style-type: circle;">
<li>Perform library amplification after the DNA isolation following the standard protocol of the chosen library preparation solution.</li>
</ul>
<h3><span>MeDIP-seq workflow</span></h3>
<center><img src="https://www.diagenode.com/img/product/kits/MeDIP-seq-workflow.png" width="110%" alt="MagMeDIP qPCR Kit x10 workflow" caption="false" /></center>
<h3><span>Example of results</span></h3>
<center><img src="https://www.diagenode.com/img/product/kits/medip-specificity.png" alt="MagMeDIP qPCR Kit Result" caption="false" width="951" height="488" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 1. qPCR analysis of external spike-in DNA controls (methylated and unmethylated) after IP.</strong> Samples were prepared using 1&mu;g &ndash; 100ng -10ng sheared human gDNA with the MagMeDIP kit (Diagenode) and a commercially available library prep kit. DNA isolation after IP has been performed with IPure kit V2 (Diagenode).</p>
<p></p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/medip-saturation-analysis.png" alt=" MagMeDIP kit " caption="false" width="951" height="461" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 2. Saturation analysis.</strong> Clean reads were aligned to the human genome (hg19) using Burrows-Wheeler aligner (BWA) algorithm after which duplicated and unmapped reads were removed resulting in a mapping efficiency &gt;98% for all samples. Quality and validity check of the mapped MeDIP-seq data was performed using MEDIPS R package. Saturation plots show that all sets of reads have sufficient complexity and depth to saturate the coverage profile of the reference genome and that this is reproducible between replicates and repetitive experiments (data shown for 50 ng gDNA input: left panel = replicate a, right panel = replicate b).</p>
<p></p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/medip-libraries-prep.png" alt="MagMeDIP x10 " caption="false" width="951" height="708" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 3. Sequencing profiles of MeDIP-seq libraries prepared from different starting amounts of sheared gDNA on the positive and negative methylated control regions.</strong> MeDIP-seq libraries were prepared from decreasing starting amounts of gDNA (1 &mu;g (green), 50 ng (red), and 10ng (blue)) originating from human blood with the MagMeDIP kit (Diagenode) and a commercially available library prep kit. DNA isolation after IP has been performed with IPure kit V2 (Diagenode). IP and corresponding INPUT samples were sequenced on Illumina NovaSeq SP with 2x50 PE reads. The reads were mapped to the human genome (hg19) with bwa and the alignments were loaded into IGV (the tracks use an identical scale). The top IGV figure shows the TSH2B (also known as H2BC1) gene (marked by blue boxes in the bottom track) and its surroundings. The TSH2B gene is coding for a histone variant that does not occur in blood cells, and it is known to be silenced by methylation. Accordingly, we see a high coverage in the vicinity of this gene. The bottom IGV figure shows the GADPH locus (marked by blue boxes in the bottom track) and its surroundings. The GADPH gene is a highly active transcription region and should not be methylated, resulting in no reads accumulation following MeDIP-seq experiment.</p>
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			'meta_description' => 'Perform Methylated DNA Immunoprecipitation (MeDIP) to estimate DNA methylation status of your sample using highly specific 5-mC antibody. This kit allows the preparation  of cfMeDIP-seq libraries.',
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<li><strong>Differential fractionation</strong> of methylated DNA by CpG density (3 eluted fractions)</li>
<li><strong>On-day protocol</strong></li>
<li><strong>NGS compatibility</strong></li>
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<h3>MBD-seq allows for detection of genomic regions with different CpG density</h3>
<p><img src="https://www.diagenode.com/img/product/kits/mbd_results1.png" alt="MBD-sequencing results have been validated by bisulfite sequencing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong></strong></p>
<p><strong></strong><strong>F</strong><strong>igure 1.</strong> Using the MBD approach, two methylated regions were detected in different elution fractions according to their methylated CpG density (A). Low, Medium and High refer to the sequenced DNA from different elution fractions with increasing salt concentration. Methylated patterns of these two different methylated regions were validated by bisulfite conversion assay (B).</p>',
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			'label1' => ' Characteristics',
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<li><strong>Fast &amp; sensitive capture</strong> of methylated DNA</li>
<li><strong>High capture efficiency</strong></li>
<li><strong>Differential fractionation</strong> of methylated DNA by CpG density (3 eluted fractions)</li>
<li><strong>Automation compatibility</strong><strong></strong>
<h3>MBD-seq allows for detection of genomic regions with different CpG density</h3>
<p><img src="https://www.diagenode.com/img/product/kits/mbd_results1.png" alt="MBD-sequencing results have been validated by bisulfite sequencing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>F</strong><strong>igure 1.</strong><span>&nbsp;</span>Using the MBD approach, two methylated regions were detected in different elution fractions according to their methylated CpG density (A). Low, Medium and High refer to the sequenced DNA from different elution fractions with increasing salt concentration. Methylated patterns of these two different methylated regions were validated by bisulfite conversion assay (B).<br /><strong></strong></p>
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<p style="text-align: center;"><strong>Make your Bisulfite conversion now in only 60 minutes !</strong></p>
<p>Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion reagent is added directly to DNA, requires no intermediate steps, and results in high yields of DNA ready for downstream analysis methods including PCR and Next-Generation Sequencing.</p>',
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			'description' => '<p><strong>Immunofluorescence</strong>:</p>
<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&deg;C, permeabilized with acetone at -20&deg;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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			'name' => 'DNA modifications',
			'description' => '<p><span style="font-weight: 400;">T</span><span style="font-weight: 400;">he pattern of <strong>DNA modifications</strong> is critical for genome stability and the control of gene expression in the cell. Methylation of 5-cytosine (5-mC), one of the best-studied epigenetic marks, is carried out by the <strong>DNA methyltransferases</strong> DNMT3A and B and DNMT1. DNMT3A and DNMT3B are responsible for </span><i><span style="font-weight: 400;">de novo</span></i><span style="font-weight: 400;"> DNA methylation, whereas DNMT1 maintains existing methylation. 5-mC undergoes active demethylation which is performed by the <strong>Ten-Eleven Translocation</strong> (TET) familly of DNA hydroxylases. The latter consists of 3 members TET1, 2 and 3. All 3 members catalyze the conversion of <strong>5-methylcytosine</strong> (5-mC) into <strong>5-hydroxymethylcytosine</strong> (5-hmC), and further into <strong>5-formylcytosine</strong> (5-fC) and <strong>5-carboxycytosine</strong> (5-caC). 5-fC and 5-caC can be converted to unmodified cytosine by <strong>Thymine DNA Glycosylase</strong> (TDG). It is not yet clear if 5-hmC, 5-fC and 5-caC have specific functions or are simply intermediates in the demethylation of 5-mC.</span></p>
<p><span style="font-weight: 400;">DNA methylation is generally considered as a repressive mark and is usually associated with gene silencing. It is essential that the balance between DNA methylation and demethylation is precisely maintained. Dysregulation of DNA methylation may lead to many different human diseases and is often observed in cancer cells.</span></p>
<p><span style="font-weight: 400;">Diagenode offers highly validated antibodies against different proteins involved in DNA modifications as well as against the modified bases allowing the study of all steps and intermediates in the DNA methylation/demethylation pathway:</span></p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/dna-methylation.jpg" height="599" width="816" /></p>
<p><strong>Diagenode exclusively sources the original 5-methylcytosine monoclonal antibody (clone 33D3).</strong></p>
<p>Check out the list below to see all proposed antibodies for DNA modifications.</p>
<p>Diagenode&rsquo;s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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<p><span style="font-weight: 400;">Diagenode&rsquo;s highly validated antibodies:</span></p>
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</form><a class="close-reveal-modal" aria-label="Close">&#215;</a></div><!-- END: ADD TO CART MODAL --><a href="#" id="auto-hmedip-kit-x16-monoclonal-mouse-antibody-16-rxns" data-reveal-id="cartModal-1885" class="" style="color:#B21329"><i class="fa fa-cart-plus"></i></a>
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<li>
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</li>
<li>
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                         <a href="/en/p/methylcap-kit-x48-48-rxns"><img src="/img/product/kits/methyl-kit-icon.png" alt="Methylation kit icon" class="th"/></a>        </div>



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            <h6 style="height:60px">MethylCap kit</h6>
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<li>
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                         <a href="/en/p/auto-methylcap-kit-x48-48-rxns"><img src="/img/product/kits/methyl-kit-icon.png" alt="Methylation kit icon" class="th"/></a>        </div>



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            <div class="small-6 columns" style="padding-left:0px;padding-right:0px;margin-top:-6px;margin-left:-1px">
                <span class="success label" style="">C02020011</span>
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        <div class="small-12 columns" >
            <h6 style="height:60px">Auto MethylCap kit</h6>
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</li>
<li>
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                    'C02030030',
                     '240',
                       $('#CartQuantity').val());" name="checkout" id="checkout" value="checkout" type="submit">Checkout</button>            </div>
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                <button class="alert small button expand" onclick="$(this).addToCart('Premium Bisulfite kit',
                    'C02030030',
                    '240',
                       $('#CartQuantity').val());" name="keepshop" id="keepshop" type="submit">Keep shopping</button>            </div>
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    </div>
</div>

</form><a class="close-reveal-modal" aria-label="Close">&#215;</a></div><!-- END: ADD TO CART MODAL --><a href="#" id="premium-bisulfite-kit-50-rxns" data-reveal-id="cartModal-1892" class="" style="color:#B21329"><i class="fa fa-cart-plus"></i></a>
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            <h6 style="height:60px">Premium Bisulfite kit</h6>
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'
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	'id' => '1892',
	'antibody_id' => null,
	'name' => 'Premium Bisulfite kit',
	'description' => '<p style="text-align: center;"><a href="https://www.diagenode.com/files/products/kits/Premium_Bisulfite_kit_manual.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p style="text-align: center;"><strong>Make your Bisulfite conversion now in only 60 minutes !</strong></p>
<p>Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion reagent is added directly to DNA, requires no intermediate steps, and results in high yields of DNA ready for downstream analysis methods including PCR and Next-Generation Sequencing.</p>',
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	'meta_description' => 'Premium Bisulfite kit',
	'modified' => '2023-04-20 16:13:50',
	'created' => '2015-06-29 14:08:20',
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)
$labelize = object(Closure) {
	
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	'id' => '3138',
	'antibody_id' => '612',
	'name' => '5-methylcytosine (5-mC) Antibody for ICC/IF (sample size)',
	'description' => '',
	'label1' => '',
	'info1' => '',
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	'info2' => '',
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	'info3' => '',
	'format' => '10 μg',
	'catalog_number' => 'C15200003-10',
	'old_catalog_number' => '',
	'sf_code' => 'C15200003-D001-000582',
	'type' => 'FRE',
	'search_order' => '03-Antibody',
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	'online' => true,
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	'last_datasheet_update' => 'July 30, 2020',
	'slug' => '5-mc-monoclonal-antibody-for-icc-if-classic-10',
	'meta_title' => '',
	'meta_keywords' => '',
	'meta_description' => '',
	'modified' => '2020-11-19 10:47:27',
	'created' => '2020-11-18 16:06:47',
	'ProductsGroup' => array(
		'id' => '357',
		'product_id' => '3138',
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$label = '<img src="/img/banners/banner-customizer-back.png" alt=""/>'
$application = array(
	'id' => '35',
	'position' => '10',
	'parent_id' => '40',
	'name' => 'FISH',
	'description' => '<p>FISH</p>',
	'in_footer' => false,
	'in_menu' => false,
	'online' => true,
	'tabular' => true,
	'slug' => 'fish-5-mC',
	'meta_keywords' => '5-Methylcytosine,5-mC,monoclonal antibody',
	'meta_description' => '5-Methylcytosine(5-mC) monoclonal antibody for fish ',
	'meta_title' => '5-Methylcytosine(5-mC) monoclonal antibody for fish | Diagenode',
	'modified' => '2016-01-13 12:24:55',
	'created' => '2015-07-08 13:53:27',
	'ProductsApplication' => array(
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)
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	'id' => '35',
	'position' => '10',
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	'name' => 'FISH',
	'description' => '<p>FISH</p>',
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	'in_menu' => false,
	'online' => true,
	'tabular' => true,
	'slug' => 'fish-5-mC',
	'meta_keywords' => '5-Methylcytosine,5-mC,monoclonal antibody',
	'meta_description' => '5-Methylcytosine(5-mC) monoclonal antibody for fish ',
	'meta_title' => '5-Methylcytosine(5-mC) monoclonal antibody for fish | Diagenode',
	'modified' => '2016-01-13 12:24:55',
	'created' => '2015-07-08 13:53:27',
	'locale' => 'eng'
)
$description = '<p>FISH</p>'
$name = 'FISH'
$document = array(
	'id' => '38',
	'name' => 'Epigenetic Antibodies Brochure',
	'description' => '<p>More than in any other immuoprecipitation assays, quality antibodies are critical tools in many epigenetics experiments. Since 10 years, Diagenode has developed the most stringent quality production available on the market for antibodies exclusively focused on epigenetic uses. All our antibodies have been qualified to work in epigenetic applications.</p>',
	'image_id' => null,
	'type' => 'Brochure',
	'url' => 'files/brochures/Epigenetic_Antibodies_Brochure.pdf',
	'slug' => 'epigenetic-antibodies-brochure',
	'meta_keywords' => '',
	'meta_description' => '',
	'modified' => '2016-06-15 11:24:06',
	'created' => '2015-07-03 16:05:27',
	'ProductsDocument' => array(
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		'product_id' => '1961',
		'document_id' => '38'
	)
)
$sds = array(
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	'name' => '5-mC antibody for ICC/IF SDS FR fr',
	'language' => 'fr',
	'url' => 'files/SDS/5-mC/SDS-C15200003-5-methylcytosine_5-mC_Antibody_for_ICC_IF-FR-fr-GHS_2_0.pdf',
	'countries' => 'FR',
	'modified' => '2021-08-24 16:32:39',
	'created' => '2021-08-24 16:32:39',
	'ProductsSafetySheet' => array(
		'id' => '2080',
		'product_id' => '1961',
		'safety_sheet_id' => '1189'
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)
$publication = array(
	'id' => '3750',
	'name' => 'RNA biochemistry. Transcriptome-wide distribution and function of RNA hydroxymethylcytosine.',
	'authors' => 'Delatte B, Wang F, Ngoc LV, Collignon E, Bonvin E, Deplus R, Calonne E, Hassabi B, Putmans P, Awe S, Wetzel C, Kreher J, Soin R, Creppe C, Limbach PA, Gueydan C, Kruys V, Brehm A, Minakhina S, Defrance M, Steward R, Fuks F',
	'description' => '<p>Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.</p>',
	'date' => '2016-01-15',
	'pmid' => 'http://www.pubmed.org/26816380',
	'doi' => '10.1126/science.aac5253',
	'modified' => '2019-10-03 12:29:05',
	'created' => '2019-10-02 16:16:55',
	'ProductsPublication' => array(
		'id' => '4128',
		'product_id' => '1961',
		'publication_id' => '3750'
	)
)
$externalLink = ' <a href="http://www.pubmed.org/26816380" target="_blank"><i class="fa fa-external-link"></i></a>'
include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118

Notice (8): Undefined variable: message [APP/View/Products/view.ctp, line 755]
Code Context
<!-- BEGIN: REQUEST_FORM MODAL -->
<div id="request_formModal" class="reveal-modal medium" data-reveal aria-labelledby="modalTitle" aria-hidden="true" role="dialog">
    <?= $this->element('Forms/simple_form', array('solution_of_interest' => $solution_of_interest, 'header' => $header, 'message' => $message, 'campaign_id' => $campaign_id)) ?>
$viewFile = '/home/website-server/www/app/View/Products/view.ctp'
$dataForView = array(
	'language' => 'en',
	'meta_keywords' => '5-methylcytosine (5-mC),monoclonal antibody ',
	'meta_description' => ' 5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF  validated  in MeDIP, DB, IF, and FISH. Batch-specific data available on the website. ',
	'meta_title' => '5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF | Diagenode',
	'product' => array(
		'Product' => array(
			'id' => '1961',
			'antibody_id' => '30',
			'name' => ' 5-methylcytosine (5-mC) Antibody for ICC/IF ',
			'description' => '<p><span>Monoclonal antibody raised in mouse against 5-mC (5-methylcytosine) conjugated to BSA.&nbsp;</span></p>',
			'label1' => 'Validation Data',
			'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_DotBlot.png" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-1.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH_2.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </small></p>
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<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
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<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </small></p>
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<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
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<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
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			'price_AUD' => '950',
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			'slug' => '5-mc-monoclonal-antibody-for-icc-if-classic-50-mg-21-ml',
			'meta_title' => '5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF | Diagenode',
			'meta_keywords' => '5-methylcytosine (5-mC),monoclonal antibody ',
			'meta_description' => ' 5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF  validated  in MeDIP, DB, IF, and FISH. Batch-specific data available on the website. ',
			'modified' => '2022-01-05 15:26:15',
			'created' => '2015-06-29 14:08:20'
		),
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			'id' => '1882',
			'antibody_id' => null,
			'name' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'description' => '<p><a href="https://www.diagenode.com/files/products/kits/hMeDIP_kit_manual.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p><span>The hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented&nbsp;genomic DNA<span><span>&nbsp;</span>samples for use in genome-wide methylation analysis. It features</span></span><span>&nbsp;a highly specific monoclonal antibody against&nbsp;</span>5-hydroxymethylcytosine (5-hmC) for the immunoprecipitation of hydroxymethylated DNA<span>. It includes&nbsp;control DNA and primers to assess the effiency of the assay.&nbsp;</span>Performing hydroxymethylation profiling with the hMeDIP kit is fast,&nbsp;reliable and&nbsp;highly specific.</p>
<p><em>Looking for hMeDIP-seq protocol? <a href="https://go.diagenode.com/l/928883/2022-01-07/2m1ht" target="_blank" title="Contact us">Contact us</a></em></p>
<p><span></span></p>
<p><span></span></p>',
			'label1' => 'Characteristics',
			'info1' => '<ul style="list-style-type: disc;">
<li><span>Robust enrichment &amp; immunoprecipitation of hydroxymethylated DNA</span></li>
<li>Highly specific monoclonal antibody against 5-hmC<span>&nbsp;for reliable, reproducible results</span></li>
<li>Including&nbsp;control DNA and primers to&nbsp;<span>monitor the&nbsp;efficiency of the assay</span>
<ul style="list-style-type: circle;">
<li>hmeDNA and unmethylated DNA sequences and primer pairs</li>
<li>Mouse primer pairs against Sfi1 targeting hydroxymethylated gene in mouse</li>
</ul>
</li>
<li>Improved single-tube, magnetic bead-based protocol</li>
</ul>',
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			'format' => '16 rxns',
			'catalog_number' => 'C02010031',
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			'sf_code' => 'C02010031-',
			'type' => 'RFR',
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			'price_EUR' => '630',
			'price_USD' => '690',
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			'slug' => 'hmedip-kit-x16-monoclonal-mouse-antibody-16-rxns',
			'meta_title' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'meta_keywords' => '',
			'meta_description' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'modified' => '2023-04-20 16:12:48',
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			'antibody_id' => null,
			'name' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'description' => '<p><span>The Auro hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented&nbsp;genomic DNA&nbsp;samples for use in genome-wide methylation analysis. It features</span><span>&nbsp;a highly specific monoclonal antibody against&nbsp;</span><span>5-hydroxymethylcytosine (5-hmC) for the immunoprecipitation of hydroxymethylated DNA</span><span>. It includes&nbsp;control DNA and primers to assess the effiency of the assay.&nbsp;</span><span>Performing hydroxymethylation profiling with the hMeDIP kit is fast,&nbsp;reliable and&nbsp;highly specific.</span></p>',
			'label1' => ' Characteristics',
			'info1' => '<ul style="list-style-type: disc;">
<li><span>Robust enrichment &amp; immunoprecipitation of hydroxymethylated DNA</span></li>
<li>Highly specific monoclonal antibody against 5-hmC<span>&nbsp;for reliable, reproducible results</span></li>
<li>Including&nbsp;control DNA and primers to&nbsp;<span>monitor the&nbsp;efficiency of the assay</span>
<ul style="list-style-type: circle;">
<li>5-hmC, 5-mC and unmethylated DNA sequences and primer pairs</li>
<li>Mouse primer pairs against Sfi1 targeting hydroxymethylated gene in mouse</li>
</ul>
</li>
</ul>
<ul style="list-style-type: disc;">
<li>Improved single-tube, magnetic bead-based protocol</li>
</ul>',
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			'format' => '16 rxns',
			'catalog_number' => 'C02010034',
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			'sf_code' => 'C02010034-',
			'type' => 'RFR',
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			'price_EUR' => '630',
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			'slug' => 'auto-hmedip-kit-x16-monoclonal-mouse-antibody-16-rxns',
			'meta_title' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'meta_keywords' => '',
			'meta_description' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'modified' => '2021-01-18 10:37:19',
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			'id' => '1880',
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			'name' => 'MagMeDIP Kit',
			'description' => '<p><a href="https://www.diagenode.com/files/products/kits/magmedip-kit-manual-C02010020-21.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>Perform <strong>MeDIP</strong> (<strong>Me</strong>thylated <strong>D</strong>NA <strong>I</strong>mmuno<strong>p</strong>recipitation) followed by qPCR or NGS to estimate DNA methylation status of your sample using a highly sensitive 5-methylcytosine antibody. Our MagMeDIP kit contains high quality reagents to get the highest enrichment of methylated DNA with an optimized user-friendly protocol.</p>
<h3><span>Features</span></h3>
<ul>
<li>Starting DNA amount: <strong>10 ng &ndash; 1 &micro;g</strong></li>
<li>Content: <strong>all reagents included</strong> for DNA extraction, immunoprecipitation (including the 5-mC antibody, spike-in controls and their corresponding qPCR primer pairs) as well as DNA isolation after IP.</li>
<li>Application: <strong>qPCR</strong> and <strong>NGS</strong></li>
<li>Robust method, <strong>superior enrichment</strong>, and easy-to-use protocol</li>
<li><strong>High reproducibility</strong> between replicates and repetitive experiments</li>
<li>Compatible with <strong>all species&nbsp;</strong></li>
</ul>
<p>&nbsp;</p>
<div class="small-12 medium-4 large-4 columns"><center></center><center></center><center></center><center><a href="https://www.ncbi.nlm.nih.gov/pubmed/30429608" target="_blank"><img src="https://www.diagenode.com/img/banners/banner-nature-publication-580.png" alt="Click here to read more about MeDIP " caption="false" width="80%" /></a></center></div>
<div class="small-12 medium-8 large-8 columns">
<h3 style="text-align: justify;">Sensitive tumour detection and classification using plasma cell-free DNA methylomes<br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/30429608" target="_blank">Read the publication</a></h3>
<h3 class="c-article-title u-h1" data-test="article-title" itemprop="name headline" style="text-align: justify;">Preparation of cfMeDIP-seq libraries for methylome profiling of plasma cell-free DNA<br /><a href="https://www.nature.com/articles/s41596-019-0202-2" target="_blank" title="cfMeDIP-seq Nature Method">Read the method</a></h3>
</div>
<p></p>
<p></p>
<p></p>
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<h3></h3>',
			'label1' => 'MagMeDIP workflow',
			'info1' => '<p>DNA methylation occurs primarily as 5-methylcytosine (5-mC), and the Diagenode MagMeDIP Kit takes advantage of a specific antibody targeting this 5-mC to immunoprecipitate methylated DNA, which can be thereafter directly analyzed by qPCR or Next-Generation Sequencing (NGS).</p>
<h3><span>How it works</span></h3>
<p>In brief, after the cell collection and lysis, the genomic DNA is extracted, sheared, and then denatured. In the next step the antibody directed against 5 methylcytosine and antibody binding beads are used for immunoselection and immunoprecipitation of methylated DNA fragments. Then, the IP&rsquo;d methylated DNA is isolated and can be used for any subsequent analysis as qPCR, amplification, hybridization on microarrays or next generation sequencing.</p>
<center><img src="https://www.diagenode.com/img/product/kits/MagMeDIP-workflow.png" width="70%" alt="5-methylcytosine" caption="false" /></center>
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			'label2' => 'MeDIP-qPCR',
			'info2' => '<p>The kit MagMeDIP contains all reagents necessary for a complete MeDIP-qPCR workflow. Two MagMeDIP protocols have been validated: for manual processing as well as for automated processing, using the Diagenode&rsquo;s IP-Star Compact Automated System (please refer to the kit manual).</p>
<ul>
<li><strong>Complete kit</strong> including DNA extraction module, IP antibody and reagents, DNA isolation buffer</li>
<li><strong>Quality control of the IP:</strong> due to methylated and unmethylated DNA spike-in controls and their associated qPCR primers</li>
<li><strong>Easy to use</strong> with user-friendly magnetic beads and rack</li>
<li><strong>Highly validated protocol</strong></li>
<li>Automated protocol supplied</li>
</ul>
<center><img src="https://www.diagenode.com/img/product/kits/fig1-magmedipkit.png" width="85%" alt="Methylated DNA Immunoprecipitation" caption="false" /></center>
<p style="font-size: 0.9em;"><em><strong>Figure 1.</strong> Immunoprecipitation results obtained with Diagenode MagMeDIP Kit</em></p>
<p style="font-size: 0.9em;">MeDIP assays were performed manually using 1 &micro;g or 50 ng gDNA from blood cells with the MagMeDIP kit (Diagenode). The IP was performed with the Methylated and Unmethylated spike-in controls included in the kit, together with the human DNA samples. The DNA was isolated/purified using DIB. Afterwards, qPCR was performed using the primer pairs included in this kit.</p>
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			'label3' => 'MeDIP-seq',
			'info3' => '<p>For DNA methylation analysis on the whole genome, MagMeDIP kit can be coupled with Next-Generation Sequencing. To perform MeDIP-sequencing we recommend the following strategy:</p>
<ul style="list-style-type: circle;">
<li>Choose a library preparation solution which is compatible with the starting amount of DNA you are planning to use (from 10 ng to 1 &mu;g). It can be a home-made solution or a commercial one.</li>
<li>Choose the indexing system that fits your needs considering the following features:</li>
<ul>
<ul>
<ul>
<li>Single-indexing, combinatorial dual-indexing or unique dual-indexing</li>
<li>Number of barcodes</li>
<li>Full-length adaptors containing the barcodes or barcoding at the final amplification step</li>
<li>Presence / absence of Unique Molecular Identifiers (for PCR duplicates removal)</li>
</ul>
</ul>
</ul>
<li>Standard library preparation protocols are compatible with double-stranded DNA only, therefore the first steps of the library preparation (end repair, A-tailing, adaptor ligation and clean-up) will have to be performed on sheared DNA, before the IP.</li>
</ul>
<p style="padding-left: 30px;"><strong>CAUTION:</strong> As the immunoprecipitation step occurs at the middle of the library preparation workflow, single-tube solutions for library preparation are usually not compatible with MeDIP-sequencing.</p>
<ul style="list-style-type: circle;">
<li>For DNA isolation after the IP, we recommend using the <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24" title="IPure kit v2">IPure kit v2</a> (available separately, Cat. No. C03010014) instead of DNA isolation Buffer.</li>
</ul>
<ul style="list-style-type: circle;">
<li>Perform library amplification after the DNA isolation following the standard protocol of the chosen library preparation solution.</li>
</ul>
<h3><span>MeDIP-seq workflow</span></h3>
<center><img src="https://www.diagenode.com/img/product/kits/MeDIP-seq-workflow.png" width="110%" alt="MagMeDIP qPCR Kit x10 workflow" caption="false" /></center>
<h3><span>Example of results</span></h3>
<center><img src="https://www.diagenode.com/img/product/kits/medip-specificity.png" alt="MagMeDIP qPCR Kit Result" caption="false" width="951" height="488" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 1. qPCR analysis of external spike-in DNA controls (methylated and unmethylated) after IP.</strong> Samples were prepared using 1&mu;g &ndash; 100ng -10ng sheared human gDNA with the MagMeDIP kit (Diagenode) and a commercially available library prep kit. DNA isolation after IP has been performed with IPure kit V2 (Diagenode).</p>
<p></p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/medip-saturation-analysis.png" alt=" MagMeDIP kit " caption="false" width="951" height="461" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 2. Saturation analysis.</strong> Clean reads were aligned to the human genome (hg19) using Burrows-Wheeler aligner (BWA) algorithm after which duplicated and unmapped reads were removed resulting in a mapping efficiency &gt;98% for all samples. Quality and validity check of the mapped MeDIP-seq data was performed using MEDIPS R package. Saturation plots show that all sets of reads have sufficient complexity and depth to saturate the coverage profile of the reference genome and that this is reproducible between replicates and repetitive experiments (data shown for 50 ng gDNA input: left panel = replicate a, right panel = replicate b).</p>
<p></p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/medip-libraries-prep.png" alt="MagMeDIP x10 " caption="false" width="951" height="708" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 3. Sequencing profiles of MeDIP-seq libraries prepared from different starting amounts of sheared gDNA on the positive and negative methylated control regions.</strong> MeDIP-seq libraries were prepared from decreasing starting amounts of gDNA (1 &mu;g (green), 50 ng (red), and 10ng (blue)) originating from human blood with the MagMeDIP kit (Diagenode) and a commercially available library prep kit. DNA isolation after IP has been performed with IPure kit V2 (Diagenode). IP and corresponding INPUT samples were sequenced on Illumina NovaSeq SP with 2x50 PE reads. The reads were mapped to the human genome (hg19) with bwa and the alignments were loaded into IGV (the tracks use an identical scale). The top IGV figure shows the TSH2B (also known as H2BC1) gene (marked by blue boxes in the bottom track) and its surroundings. The TSH2B gene is coding for a histone variant that does not occur in blood cells, and it is known to be silenced by methylation. Accordingly, we see a high coverage in the vicinity of this gene. The bottom IGV figure shows the GADPH locus (marked by blue boxes in the bottom track) and its surroundings. The GADPH gene is a highly active transcription region and should not be methylated, resulting in no reads accumulation following MeDIP-seq experiment.</p>
<p></p>
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			'slug' => 'magmedip-kit-x48-48-rxns',
			'meta_title' => 'MagMeDIP Kit for efficient immunoprecipitation of methylated DNA | Diagenode',
			'meta_keywords' => '',
			'meta_description' => 'Perform Methylated DNA Immunoprecipitation (MeDIP) to estimate DNA methylation status of your sample using highly specific 5-mC antibody. This kit allows the preparation  of cfMeDIP-seq libraries.',
			'modified' => '2024-12-04 16:52:47',
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		(int) 3 => array(
			'id' => '1887',
			'antibody_id' => null,
			'name' => 'MethylCap kit',
			'description' => '<p>The MethylCap kit allows to specifically capture DNA fragments containing methylated CpGs. The assay is based on the affinity purification of methylated DNA using methyl-CpG-binding domain (MBD) of human MeCP2 protein. The procedure has been adapted to both manual process or <a href="https://www.diagenode.com/en/p/sx-8g-ip-star-compact-automated-system-1-unit">IP-Star&reg; Compact Automated System</a>. Libraries of captured methylated DNA can be prepared for next-generation sequencing (NGS) by combining MBD technology with the <a href="https://www.diagenode.com/en/p/microplex-lib-prep-kit-v3-48-rxns">MicroPlex Library Preparation Kit v3</a>.</p>',
			'label1' => 'Characteristics',
			'info1' => '<ul style="list-style-type: circle;">
<li><strong>Fast &amp; sensitive capture</strong> of methylated DNA</li>
<li><strong>High capture efficiency</strong></li>
<li><strong>Differential fractionation</strong> of methylated DNA by CpG density (3 eluted fractions)</li>
<li><strong>On-day protocol</strong></li>
<li><strong>NGS compatibility</strong></li>
</ul>
<h3>MBD-seq allows for detection of genomic regions with different CpG density</h3>
<p><img src="https://www.diagenode.com/img/product/kits/mbd_results1.png" alt="MBD-sequencing results have been validated by bisulfite sequencing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong></strong></p>
<p><strong></strong><strong>F</strong><strong>igure 1.</strong> Using the MBD approach, two methylated regions were detected in different elution fractions according to their methylated CpG density (A). Low, Medium and High refer to the sequenced DNA from different elution fractions with increasing salt concentration. Methylated patterns of these two different methylated regions were validated by bisulfite conversion assay (B).</p>',
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			'slug' => 'methylcap-kit-x48-48-rxns',
			'meta_title' => 'MethylCap kit x48',
			'meta_keywords' => '',
			'meta_description' => 'MethylCap kit x48',
			'modified' => '2024-11-21 06:38:46',
			'created' => '2015-06-29 14:08:20',
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			'id' => '1888',
			'antibody_id' => null,
			'name' => 'Auto MethylCap kit',
			'description' => '<p>The Auto MethylCap kit allows to specifically capture DNA fragments containing methylated CpGs. The assay is based on the affinity purification of methylated DNA using methyl-CpG-binding domain (MBD) of human MeCP2 protein. This procedure has been optimized to perform automated immunoprecipitation of chromatin using the <a href="https://www.diagenode.com/en/p/sx-8g-ip-star-compact-automated-system-1-unit">IP-Star&reg; Compact Automated System</a> enabling highly reproducible results and allowing for high throughput.</p>',
			'label1' => ' Characteristics',
			'info1' => '<ul>
<li><strong>Fast &amp; sensitive capture</strong> of methylated DNA</li>
<li><strong>High capture efficiency</strong></li>
<li><strong>Differential fractionation</strong> of methylated DNA by CpG density (3 eluted fractions)</li>
<li><strong>Automation compatibility</strong><strong></strong>
<h3>MBD-seq allows for detection of genomic regions with different CpG density</h3>
<p><img src="https://www.diagenode.com/img/product/kits/mbd_results1.png" alt="MBD-sequencing results have been validated by bisulfite sequencing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>F</strong><strong>igure 1.</strong><span>&nbsp;</span>Using the MBD approach, two methylated regions were detected in different elution fractions according to their methylated CpG density (A). Low, Medium and High refer to the sequenced DNA from different elution fractions with increasing salt concentration. Methylated patterns of these two different methylated regions were validated by bisulfite conversion assay (B).<br /><strong></strong></p>
</li>
</ul>',
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<p style="text-align: center;"><strong>Make your Bisulfite conversion now in only 60 minutes !</strong></p>
<p>Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion reagent is added directly to DNA, requires no intermediate steps, and results in high yields of DNA ready for downstream analysis methods including PCR and Next-Generation Sequencing.</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&deg;C, permeabilized with acetone at -20&deg;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><span style="font-weight: 400;">T</span><span style="font-weight: 400;">he pattern of <strong>DNA modifications</strong> is critical for genome stability and the control of gene expression in the cell. Methylation of 5-cytosine (5-mC), one of the best-studied epigenetic marks, is carried out by the <strong>DNA methyltransferases</strong> DNMT3A and B and DNMT1. DNMT3A and DNMT3B are responsible for </span><i><span style="font-weight: 400;">de novo</span></i><span style="font-weight: 400;"> DNA methylation, whereas DNMT1 maintains existing methylation. 5-mC undergoes active demethylation which is performed by the <strong>Ten-Eleven Translocation</strong> (TET) familly of DNA hydroxylases. The latter consists of 3 members TET1, 2 and 3. All 3 members catalyze the conversion of <strong>5-methylcytosine</strong> (5-mC) into <strong>5-hydroxymethylcytosine</strong> (5-hmC), and further into <strong>5-formylcytosine</strong> (5-fC) and <strong>5-carboxycytosine</strong> (5-caC). 5-fC and 5-caC can be converted to unmodified cytosine by <strong>Thymine DNA Glycosylase</strong> (TDG). It is not yet clear if 5-hmC, 5-fC and 5-caC have specific functions or are simply intermediates in the demethylation of 5-mC.</span></p>
<p><span style="font-weight: 400;">DNA methylation is generally considered as a repressive mark and is usually associated with gene silencing. It is essential that the balance between DNA methylation and demethylation is precisely maintained. Dysregulation of DNA methylation may lead to many different human diseases and is often observed in cancer cells.</span></p>
<p><span style="font-weight: 400;">Diagenode offers highly validated antibodies against different proteins involved in DNA modifications as well as against the modified bases allowing the study of all steps and intermediates in the DNA methylation/demethylation pathway:</span></p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/dna-methylation.jpg" height="599" width="816" /></p>
<p><strong>Diagenode exclusively sources the original 5-methylcytosine monoclonal antibody (clone 33D3).</strong></p>
<p>Check out the list below to see all proposed antibodies for DNA modifications.</p>
<p>Diagenode&rsquo;s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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			'meta_description' => 'Diagenode offers Monoclonal and Polyclonal antibodies for DNA Methylation. The pattern of DNA modifications is critical for genome stability and the control of gene expression in the cell. ',
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			'name' => 'All antibodies',
			'description' => '<p><span style="font-weight: 400;">All Diagenode&rsquo;s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode&rsquo;s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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			'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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			'name' => 'Developmental mRNA mC landscape and regulatory innovations of massivemC modification of maternal mRNAs in animals.',
			'authors' => 'Liu  J. et al.',
			'description' => '<p>mC is one of the longest-known RNA modifications, however, its developmental dynamics, functions, and evolution in mRNAs remain largely unknown. Here, we generate quantitative mRNA mC maps at different stages of development in 6 vertebrate and invertebrate species and find convergent and unexpected massive methylation of maternal mRNAs mediated by NSUN2 and NSUN6. Using Drosophila as a model, we reveal that embryos lacking maternal mRNA mC undergo cell cycle delays and fail to timely initiate maternal-to-zygotic transition, implying the functional importance of maternal mRNA mC. From invertebrates to the lineage leading to humans, two waves of mC regulatory innovations are observed: higher animals gain cis-directed NSUN2-mediated mC sites at the 5' end of the mRNAs, accompanied by the emergence of more structured 5'UTR regions; humans gain thousands of trans-directed NSUN6-mediated mC sites enriched in genes regulating the mitotic cell cycle. Collectively, our studies highlight the existence and regulatory innovations of a mechanism of early embryonic development and provide key resources for elucidating the role of mRNA mC in biology and disease.</p>',
			'date' => '2022-05-01',
			'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/35513466',
			'doi' => '10.1038/s41467-022-30210-0',
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			'authors' => 'Wang, Zhen et al.',
			'description' => '<p>Accurate decoding of nucleic acid variation is critical to understand the complexity and regulation of genome function. Here we use a single-molecule magnetic tweezer (MT) platform to identify sequence variation and map a range of important epigenetic base modifications with high sensitivity, specificity, and precision in the same single molecules of DNA or RNA. We have also developed a highly specific amplification-free CRISPR-Cas enrichment strategy to isolate genomic regions from native DNA. We demonstrate enrichment of DNA from both E. coli and the FMR1 5'UTR coming from cells derived from a Fragile X carrier. From these kilobase-length enriched molecules we could characterize the differential levels of adenine and cytosine base modifications on E. coli, and the repeat expansion length and methylation status of FMR1. Together these results demonstrate that our platform can detect a variety of genetic, epigenetic, and base modification changes concomitantly within the same single molecules.</p>',
			'date' => '2021-01-01',
			'pmid' => 'https://www.ncbi.nlm.nih.gov/pubmed/33514840',
			'doi' => '10.1038/s42003-021-01648-7',
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			'authors' => 'Delatte B, Wang F, Ngoc LV, Collignon E, Bonvin E, Deplus R, Calonne E, Hassabi B, Putmans P, Awe S, Wetzel C, Kreher J, Soin R, Creppe C, Limbach PA, Gueydan C, Kruys V, Brehm A, Minakhina S, Defrance M, Steward R, Fuks F.',
			'description' => '<p>Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.</p>',
			'date' => '2016-01-15',
			'pmid' => 'http://pubmed.gov/26816380',
			'doi' => '10.1126/science.aac5253',
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			'description' => '<p>Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.</p>',
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	'url' => 'files/SDS/5-mC/SDS-C15200003-5-methylcytosine_5-mC_Antibody_for_ICC_IF-FR-fr-GHS_2_0.pdf',
	'countries' => 'FR',
	'modified' => '2021-08-24 16:32:39',
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	'name' => 'RNA biochemistry. Transcriptome-wide distribution and function of RNA hydroxymethylcytosine.',
	'authors' => 'Delatte B, Wang F, Ngoc LV, Collignon E, Bonvin E, Deplus R, Calonne E, Hassabi B, Putmans P, Awe S, Wetzel C, Kreher J, Soin R, Creppe C, Limbach PA, Gueydan C, Kruys V, Brehm A, Minakhina S, Defrance M, Steward R, Fuks F',
	'description' => '<p>Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.</p>',
	'date' => '2016-01-15',
	'pmid' => 'http://www.pubmed.org/26816380',
	'doi' => '10.1126/science.aac5253',
	'modified' => '2019-10-03 12:29:05',
	'created' => '2019-10-02 16:16:55',
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$externalLink = ' <a href="http://www.pubmed.org/26816380" target="_blank"><i class="fa fa-external-link"></i></a>'
include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118

Notice (8): Undefined variable: campaign_id [APP/View/Products/view.ctp, line 755]
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<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<th>Suggested dilution</th>
<th>References</th>
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<td>MeDIP <sup>*</sup></td>
<td>1-2 &micro;g per IP</td>
<td></td>
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<td>Dot Blotting **</td>
<td>1:600</td>
<td>Fig 1</td>
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<td>Immunofluorescence</td>
<td>1:1,000</td>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 &micro;g per IP.</small></p>
<p><small><sup>**</sup> Dot blot was only performed to demonstrate the specificity. This antibody is not recommended for dot blot on biological samples</small></p>',
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<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
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<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
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<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
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<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
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		'isotype' => 'IgG1',
		'lot' => '003',
		'concentration' => '1.3 &micro;g/&micro;l',
		'reactivity' => 'Human, mouse, drosophila, other (wide range): positive.',
		'type' => 'Monoclonal',
		'purity' => 'Protein A purified monoclonal antibody.',
		'classification' => 'Classic',
		'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>MeDIP <sup>*</sup></td>
<td>1-2 &micro;g per IP</td>
<td></td>
</tr>
<tr>
<td>Dot Blotting **</td>
<td>1:600</td>
<td>Fig 1</td>
</tr>
<tr>
<td>Immunofluorescence</td>
<td>1:1,000</td>
<td>Fig 2, 3</td>
</tr>
<tr>
<td>FISH</td>
<td>1:200 - 1:1,000</td>
<td>Fig 4, 5</td>
</tr>
</tbody>
</table>
<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 &micro;g per IP.</small></p>
<p><small><sup>**</sup> Dot blot was only performed to demonstrate the specificity. This antibody is not recommended for dot blot on biological samples</small></p>',
		'storage_conditions' => 'Store at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.',
		'storage_buffer' => 'PBS containing 0.05% azide.',
		'precautions' => 'This product is for research use only. Not for use in diagnostic or therapeutic procedures.',
		'uniprot_acc' => '',
		'slug' => '',
		'meta_keywords' => '',
		'meta_description' => '',
		'modified' => '2020-07-30 15:53:25',
		'created' => '0000-00-00 00:00:00',
		'select_label' => '30 - 5-mC monoclonal antibody for ICC/IF (003 - 1.3 &micro;g/&micro;l - Human, mouse, drosophila, other (wide range): positive. - Protein A purified monoclonal antibody. - Mouse)'
	),
	'Slave' => array(
		(int) 0 => array(
			'id' => '317',
			'name' => 'C15200003',
			'product_id' => '1961',
			'modified' => '2020-11-18 16:07:10',
			'created' => '2020-11-18 16:07:10'
		)
	),
	'Group' => array(
		'Group' => array(
			'id' => '317',
			'name' => 'C15200003',
			'product_id' => '1961',
			'modified' => '2020-11-18 16:07:10',
			'created' => '2020-11-18 16:07:10'
		),
		'Master' => array(
			'id' => '1961',
			'antibody_id' => '30',
			'name' => ' 5-methylcytosine (5-mC) Antibody for ICC/IF ',
			'description' => '<p><span>Monoclonal antibody raised in mouse against 5-mC (5-methylcytosine) conjugated to BSA.&nbsp;</span></p>',
			'label1' => 'Validation Data',
			'info1' => '<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_DotBlot.png" alt="Dot Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 1. Dot blot analysis using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To demonstrate the specificity of the Diagenode antibody against 5-mC (Cat. No. C15200003), a Dot blot analysis was performed using the hmC, mC and C controls from the Diagenode &ldquo;5-hmC, 5-mC &amp; cytosine DNA Standard Pack&rdquo; (Cat. No. AF-101-0002). One hundred to 4 ng (equivalent of 5 to 0.2 pmol of C-bases) of the controls were spotted on a membrane (Amersham Hybond-N+). The antibody was used at a dilution of 1:600. Figure 1 shows a high specificity of the antibody for the methylated control. </small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-1.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 2. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />HeLa cells were stained with the Diagenode antibody against 5-mC (Cat. No. C15200003) and with DAPI. Cells were fixed with 4% formaldehyde for 10&rsquo; and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labelled with the 5-mC antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><small>A.</small><img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2A.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><small>B.</small> <img src="https://www.diagenode.com/img/product/antibodies/C15200003-IF-2B.png" alt="Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-8 columns">
<p><small><strong>Figure 3. Immunofluorescence results obtained with the Diagenode monoclonal antibody directed against 5-mC </strong><br />Human osteosarcoma (U2OS) cells were stained with the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). Cells were fixed with 2.5% PFA in PBS for 30&rsquo;, permeabilised with 0.5% Triton X-100 for 1 hour and treated with 2N HCl for 1 hour followed by 2 x 5 minutes with 0.1 M borate buffer to depurinate the DNA. After blocking with PBS containing 0.1% Triton X-100 and 1% BSA, the cells were immunofluorescently labelled with the 5-mC antibody diluted 1:500 in blocking solution, followed by a goat anti-mouse antibody conjugated to Alexa488. Figure 3A: cells were immunofluorescently labelled with the 5-mC antibody after incubation of the antibody with 50 &micro;M mCTP (left) or with DAPI (right). Figure 3C: staining of the cells with the 5-mC antibody after incubation of the antibody with 50 &micro;M hmCTP and with DAPI.</small></p>
</div>
</div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="extra-spaced"></div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH_2.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 4. FISH using the Diagenode monoclonal antibody directed against 5-mC</strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right.</small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200003_FISH.png" alt="Fish" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 5. FISH using the Diagenode monoclonal antibody directed against 5-mC </strong><br />To detect methylated chromosomal regions, FISH was performed on metaphase chromosomes from HeLa cells using the Diagenode monoclonal antibody against 5-mC (Cat. No. C15200003). The cells were blocked in metaphase by treatment with colcemid (0.1 &micro;g/ml) for 1 - 2 hours, fixed overnight at -20&deg;C with ethanol/glacial acetic acid and treated with 2N HCl for 30&rsquo; at room temperature. Subsequently, the cells were blocked with PBS containing 1% BSA and 0.1% Triton X-100 and stained with the 5-mC antibody (left) diluted 1:1,000 in blocking solution, followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the chromosomes with DAPI. A merge of the two stainings is shown on the right. </small></p>
</div>
</div>',
			'label2' => '',
			'info2' => '',
			'label3' => '',
			'info3' => '',
			'format' => '50 μg/39 μl',
			'catalog_number' => 'C15200003',
			'old_catalog_number' => '',
			'sf_code' => 'C15200003-D001-000581',
			'type' => 'FRE',
			'search_order' => '03-Antibody',
			'price_EUR' => '380',
			'price_USD' => '380',
			'price_GBP' => '340',
			'price_JPY' => '59525',
			'price_CNY' => '',
			'price_AUD' => '950',
			'country' => 'ALL',
			'except_countries' => 'None',
			'quote' => false,
			'in_stock' => false,
			'featured' => false,
			'no_promo' => false,
			'online' => true,
			'master' => true,
			'last_datasheet_update' => 'July 30, 2020',
			'slug' => '5-mc-monoclonal-antibody-for-icc-if-classic-50-mg-21-ml',
			'meta_title' => '5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF | Diagenode',
			'meta_keywords' => '5-methylcytosine (5-mC),monoclonal antibody ',
			'meta_description' => ' 5-methylcytosine (5-mC) Monoclonal Antibody for ICC/IF  validated  in MeDIP, DB, IF, and FISH. Batch-specific data available on the website. ',
			'modified' => '2022-01-05 15:26:15',
			'created' => '2015-06-29 14:08:20'
		),
		'Product' => array(
			(int) 0 => array(
				[maximum depth reached]
			)
		)
	),
	'Related' => array(
		(int) 0 => array(
			'id' => '1882',
			'antibody_id' => null,
			'name' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'description' => '<p><a href="https://www.diagenode.com/files/products/kits/hMeDIP_kit_manual.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p><span>The hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented&nbsp;genomic DNA<span><span>&nbsp;</span>samples for use in genome-wide methylation analysis. It features</span></span><span>&nbsp;a highly specific monoclonal antibody against&nbsp;</span>5-hydroxymethylcytosine (5-hmC) for the immunoprecipitation of hydroxymethylated DNA<span>. It includes&nbsp;control DNA and primers to assess the effiency of the assay.&nbsp;</span>Performing hydroxymethylation profiling with the hMeDIP kit is fast,&nbsp;reliable and&nbsp;highly specific.</p>
<p><em>Looking for hMeDIP-seq protocol? <a href="https://go.diagenode.com/l/928883/2022-01-07/2m1ht" target="_blank" title="Contact us">Contact us</a></em></p>
<p><span></span></p>
<p><span></span></p>',
			'label1' => 'Characteristics',
			'info1' => '<ul style="list-style-type: disc;">
<li><span>Robust enrichment &amp; immunoprecipitation of hydroxymethylated DNA</span></li>
<li>Highly specific monoclonal antibody against 5-hmC<span>&nbsp;for reliable, reproducible results</span></li>
<li>Including&nbsp;control DNA and primers to&nbsp;<span>monitor the&nbsp;efficiency of the assay</span>
<ul style="list-style-type: circle;">
<li>hmeDNA and unmethylated DNA sequences and primer pairs</li>
<li>Mouse primer pairs against Sfi1 targeting hydroxymethylated gene in mouse</li>
</ul>
</li>
<li>Improved single-tube, magnetic bead-based protocol</li>
</ul>',
			'label2' => '',
			'info2' => '',
			'label3' => '',
			'info3' => '',
			'format' => '16 rxns',
			'catalog_number' => 'C02010031',
			'old_catalog_number' => 'AF-110-0016',
			'sf_code' => 'C02010031-',
			'type' => 'RFR',
			'search_order' => '04-undefined',
			'price_EUR' => '630',
			'price_USD' => '690',
			'price_GBP' => '580',
			'price_JPY' => '98690',
			'price_CNY' => '',
			'price_AUD' => '1725',
			'country' => 'ALL',
			'except_countries' => 'None',
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			'online' => true,
			'master' => true,
			'last_datasheet_update' => '0000-00-00',
			'slug' => 'hmedip-kit-x16-monoclonal-mouse-antibody-16-rxns',
			'meta_title' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'meta_keywords' => '',
			'meta_description' => 'hMeDIP kit x16 (monoclonal mouse antibody)',
			'modified' => '2023-04-20 16:12:48',
			'created' => '2015-06-29 14:08:20',
			'ProductsRelated' => array(
				[maximum depth reached]
			),
			'Image' => array(
				[maximum depth reached]
			)
		),
		(int) 1 => array(
			'id' => '1885',
			'antibody_id' => null,
			'name' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'description' => '<p><span>The Auro hMeDIP kit is designed for enrichment of hydroxymethylated DNA from fragmented&nbsp;genomic DNA&nbsp;samples for use in genome-wide methylation analysis. It features</span><span>&nbsp;a highly specific monoclonal antibody against&nbsp;</span><span>5-hydroxymethylcytosine (5-hmC) for the immunoprecipitation of hydroxymethylated DNA</span><span>. It includes&nbsp;control DNA and primers to assess the effiency of the assay.&nbsp;</span><span>Performing hydroxymethylation profiling with the hMeDIP kit is fast,&nbsp;reliable and&nbsp;highly specific.</span></p>',
			'label1' => ' Characteristics',
			'info1' => '<ul style="list-style-type: disc;">
<li><span>Robust enrichment &amp; immunoprecipitation of hydroxymethylated DNA</span></li>
<li>Highly specific monoclonal antibody against 5-hmC<span>&nbsp;for reliable, reproducible results</span></li>
<li>Including&nbsp;control DNA and primers to&nbsp;<span>monitor the&nbsp;efficiency of the assay</span>
<ul style="list-style-type: circle;">
<li>5-hmC, 5-mC and unmethylated DNA sequences and primer pairs</li>
<li>Mouse primer pairs against Sfi1 targeting hydroxymethylated gene in mouse</li>
</ul>
</li>
</ul>
<ul style="list-style-type: disc;">
<li>Improved single-tube, magnetic bead-based protocol</li>
</ul>',
			'label2' => '',
			'info2' => '',
			'label3' => '',
			'info3' => '',
			'format' => '16 rxns',
			'catalog_number' => 'C02010034',
			'old_catalog_number' => '',
			'sf_code' => 'C02010034-',
			'type' => 'RFR',
			'search_order' => '04-undefined',
			'price_EUR' => '630',
			'price_USD' => '690',
			'price_GBP' => '580',
			'price_JPY' => '98690',
			'price_CNY' => '',
			'price_AUD' => '1725',
			'country' => 'ALL',
			'except_countries' => 'Japan',
			'quote' => false,
			'in_stock' => false,
			'featured' => false,
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			'online' => true,
			'master' => true,
			'last_datasheet_update' => '0000-00-00',
			'slug' => 'auto-hmedip-kit-x16-monoclonal-mouse-antibody-16-rxns',
			'meta_title' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'meta_keywords' => '',
			'meta_description' => 'Auto hMeDIP kit x16 (monoclonal mouse antibody)',
			'modified' => '2021-01-18 10:37:19',
			'created' => '2015-06-29 14:08:20',
			'ProductsRelated' => array(
				[maximum depth reached]
			),
			'Image' => array(
				[maximum depth reached]
			)
		),
		(int) 2 => array(
			'id' => '1880',
			'antibody_id' => null,
			'name' => 'MagMeDIP Kit',
			'description' => '<p><a href="https://www.diagenode.com/files/products/kits/magmedip-kit-manual-C02010020-21.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>Perform <strong>MeDIP</strong> (<strong>Me</strong>thylated <strong>D</strong>NA <strong>I</strong>mmuno<strong>p</strong>recipitation) followed by qPCR or NGS to estimate DNA methylation status of your sample using a highly sensitive 5-methylcytosine antibody. Our MagMeDIP kit contains high quality reagents to get the highest enrichment of methylated DNA with an optimized user-friendly protocol.</p>
<h3><span>Features</span></h3>
<ul>
<li>Starting DNA amount: <strong>10 ng &ndash; 1 &micro;g</strong></li>
<li>Content: <strong>all reagents included</strong> for DNA extraction, immunoprecipitation (including the 5-mC antibody, spike-in controls and their corresponding qPCR primer pairs) as well as DNA isolation after IP.</li>
<li>Application: <strong>qPCR</strong> and <strong>NGS</strong></li>
<li>Robust method, <strong>superior enrichment</strong>, and easy-to-use protocol</li>
<li><strong>High reproducibility</strong> between replicates and repetitive experiments</li>
<li>Compatible with <strong>all species&nbsp;</strong></li>
</ul>
<p>&nbsp;</p>
<div class="small-12 medium-4 large-4 columns"><center></center><center></center><center></center><center><a href="https://www.ncbi.nlm.nih.gov/pubmed/30429608" target="_blank"><img src="https://www.diagenode.com/img/banners/banner-nature-publication-580.png" alt="Click here to read more about MeDIP " caption="false" width="80%" /></a></center></div>
<div class="small-12 medium-8 large-8 columns">
<h3 style="text-align: justify;">Sensitive tumour detection and classification using plasma cell-free DNA methylomes<br /><a href="https://www.ncbi.nlm.nih.gov/pubmed/30429608" target="_blank">Read the publication</a></h3>
<h3 class="c-article-title u-h1" data-test="article-title" itemprop="name headline" style="text-align: justify;">Preparation of cfMeDIP-seq libraries for methylome profiling of plasma cell-free DNA<br /><a href="https://www.nature.com/articles/s41596-019-0202-2" target="_blank" title="cfMeDIP-seq Nature Method">Read the method</a></h3>
</div>
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<p></p>
<p></p>
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<h3></h3>',
			'label1' => 'MagMeDIP workflow',
			'info1' => '<p>DNA methylation occurs primarily as 5-methylcytosine (5-mC), and the Diagenode MagMeDIP Kit takes advantage of a specific antibody targeting this 5-mC to immunoprecipitate methylated DNA, which can be thereafter directly analyzed by qPCR or Next-Generation Sequencing (NGS).</p>
<h3><span>How it works</span></h3>
<p>In brief, after the cell collection and lysis, the genomic DNA is extracted, sheared, and then denatured. In the next step the antibody directed against 5 methylcytosine and antibody binding beads are used for immunoselection and immunoprecipitation of methylated DNA fragments. Then, the IP&rsquo;d methylated DNA is isolated and can be used for any subsequent analysis as qPCR, amplification, hybridization on microarrays or next generation sequencing.</p>
<center><img src="https://www.diagenode.com/img/product/kits/MagMeDIP-workflow.png" width="70%" alt="5-methylcytosine" caption="false" /></center>
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<script src="chrome-extension://hhojmcideegachlhfgfdhailpfhgknjm/web_accessible_resources/index.js"></script>',
			'label2' => 'MeDIP-qPCR',
			'info2' => '<p>The kit MagMeDIP contains all reagents necessary for a complete MeDIP-qPCR workflow. Two MagMeDIP protocols have been validated: for manual processing as well as for automated processing, using the Diagenode&rsquo;s IP-Star Compact Automated System (please refer to the kit manual).</p>
<ul>
<li><strong>Complete kit</strong> including DNA extraction module, IP antibody and reagents, DNA isolation buffer</li>
<li><strong>Quality control of the IP:</strong> due to methylated and unmethylated DNA spike-in controls and their associated qPCR primers</li>
<li><strong>Easy to use</strong> with user-friendly magnetic beads and rack</li>
<li><strong>Highly validated protocol</strong></li>
<li>Automated protocol supplied</li>
</ul>
<center><img src="https://www.diagenode.com/img/product/kits/fig1-magmedipkit.png" width="85%" alt="Methylated DNA Immunoprecipitation" caption="false" /></center>
<p style="font-size: 0.9em;"><em><strong>Figure 1.</strong> Immunoprecipitation results obtained with Diagenode MagMeDIP Kit</em></p>
<p style="font-size: 0.9em;">MeDIP assays were performed manually using 1 &micro;g or 50 ng gDNA from blood cells with the MagMeDIP kit (Diagenode). The IP was performed with the Methylated and Unmethylated spike-in controls included in the kit, together with the human DNA samples. The DNA was isolated/purified using DIB. Afterwards, qPCR was performed using the primer pairs included in this kit.</p>
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			'label3' => 'MeDIP-seq',
			'info3' => '<p>For DNA methylation analysis on the whole genome, MagMeDIP kit can be coupled with Next-Generation Sequencing. To perform MeDIP-sequencing we recommend the following strategy:</p>
<ul style="list-style-type: circle;">
<li>Choose a library preparation solution which is compatible with the starting amount of DNA you are planning to use (from 10 ng to 1 &mu;g). It can be a home-made solution or a commercial one.</li>
<li>Choose the indexing system that fits your needs considering the following features:</li>
<ul>
<ul>
<ul>
<li>Single-indexing, combinatorial dual-indexing or unique dual-indexing</li>
<li>Number of barcodes</li>
<li>Full-length adaptors containing the barcodes or barcoding at the final amplification step</li>
<li>Presence / absence of Unique Molecular Identifiers (for PCR duplicates removal)</li>
</ul>
</ul>
</ul>
<li>Standard library preparation protocols are compatible with double-stranded DNA only, therefore the first steps of the library preparation (end repair, A-tailing, adaptor ligation and clean-up) will have to be performed on sheared DNA, before the IP.</li>
</ul>
<p style="padding-left: 30px;"><strong>CAUTION:</strong> As the immunoprecipitation step occurs at the middle of the library preparation workflow, single-tube solutions for library preparation are usually not compatible with MeDIP-sequencing.</p>
<ul style="list-style-type: circle;">
<li>For DNA isolation after the IP, we recommend using the <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24" title="IPure kit v2">IPure kit v2</a> (available separately, Cat. No. C03010014) instead of DNA isolation Buffer.</li>
</ul>
<ul style="list-style-type: circle;">
<li>Perform library amplification after the DNA isolation following the standard protocol of the chosen library preparation solution.</li>
</ul>
<h3><span>MeDIP-seq workflow</span></h3>
<center><img src="https://www.diagenode.com/img/product/kits/MeDIP-seq-workflow.png" width="110%" alt="MagMeDIP qPCR Kit x10 workflow" caption="false" /></center>
<h3><span>Example of results</span></h3>
<center><img src="https://www.diagenode.com/img/product/kits/medip-specificity.png" alt="MagMeDIP qPCR Kit Result" caption="false" width="951" height="488" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 1. qPCR analysis of external spike-in DNA controls (methylated and unmethylated) after IP.</strong> Samples were prepared using 1&mu;g &ndash; 100ng -10ng sheared human gDNA with the MagMeDIP kit (Diagenode) and a commercially available library prep kit. DNA isolation after IP has been performed with IPure kit V2 (Diagenode).</p>
<p></p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/medip-saturation-analysis.png" alt=" MagMeDIP kit " caption="false" width="951" height="461" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 2. Saturation analysis.</strong> Clean reads were aligned to the human genome (hg19) using Burrows-Wheeler aligner (BWA) algorithm after which duplicated and unmapped reads were removed resulting in a mapping efficiency &gt;98% for all samples. Quality and validity check of the mapped MeDIP-seq data was performed using MEDIPS R package. Saturation plots show that all sets of reads have sufficient complexity and depth to saturate the coverage profile of the reference genome and that this is reproducible between replicates and repetitive experiments (data shown for 50 ng gDNA input: left panel = replicate a, right panel = replicate b).</p>
<p></p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/medip-libraries-prep.png" alt="MagMeDIP x10 " caption="false" width="951" height="708" /></center>
<p></p>
<p style="font-size: 0.9em;"><strong>Figure 3. Sequencing profiles of MeDIP-seq libraries prepared from different starting amounts of sheared gDNA on the positive and negative methylated control regions.</strong> MeDIP-seq libraries were prepared from decreasing starting amounts of gDNA (1 &mu;g (green), 50 ng (red), and 10ng (blue)) originating from human blood with the MagMeDIP kit (Diagenode) and a commercially available library prep kit. DNA isolation after IP has been performed with IPure kit V2 (Diagenode). IP and corresponding INPUT samples were sequenced on Illumina NovaSeq SP with 2x50 PE reads. The reads were mapped to the human genome (hg19) with bwa and the alignments were loaded into IGV (the tracks use an identical scale). The top IGV figure shows the TSH2B (also known as H2BC1) gene (marked by blue boxes in the bottom track) and its surroundings. The TSH2B gene is coding for a histone variant that does not occur in blood cells, and it is known to be silenced by methylation. Accordingly, we see a high coverage in the vicinity of this gene. The bottom IGV figure shows the GADPH locus (marked by blue boxes in the bottom track) and its surroundings. The GADPH gene is a highly active transcription region and should not be methylated, resulting in no reads accumulation following MeDIP-seq experiment.</p>
<p></p>
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<h3>MBD-seq allows for detection of genomic regions with different CpG density</h3>
<p><img src="https://www.diagenode.com/img/product/kits/mbd_results1.png" alt="MBD-sequencing results have been validated by bisulfite sequencing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong></strong></p>
<p><strong></strong><strong>F</strong><strong>igure 1.</strong> Using the MBD approach, two methylated regions were detected in different elution fractions according to their methylated CpG density (A). Low, Medium and High refer to the sequenced DNA from different elution fractions with increasing salt concentration. Methylated patterns of these two different methylated regions were validated by bisulfite conversion assay (B).</p>',
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<h3>MBD-seq allows for detection of genomic regions with different CpG density</h3>
<p><img src="https://www.diagenode.com/img/product/kits/mbd_results1.png" alt="MBD-sequencing results have been validated by bisulfite sequencing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p style="text-align: center;"><strong>Make your Bisulfite conversion now in only 60 minutes !</strong></p>
<p>Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion reagent is added directly to DNA, requires no intermediate steps, and results in high yields of DNA ready for downstream analysis methods including PCR and Next-Generation Sequencing.</p>',
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			'description' => '<p><strong>Immunofluorescence</strong>:</p>
<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&deg;C, permeabilized with acetone at -20&deg;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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			'name' => 'DNA modifications',
			'description' => '<p><span style="font-weight: 400;">T</span><span style="font-weight: 400;">he pattern of <strong>DNA modifications</strong> is critical for genome stability and the control of gene expression in the cell. Methylation of 5-cytosine (5-mC), one of the best-studied epigenetic marks, is carried out by the <strong>DNA methyltransferases</strong> DNMT3A and B and DNMT1. DNMT3A and DNMT3B are responsible for </span><i><span style="font-weight: 400;">de novo</span></i><span style="font-weight: 400;"> DNA methylation, whereas DNMT1 maintains existing methylation. 5-mC undergoes active demethylation which is performed by the <strong>Ten-Eleven Translocation</strong> (TET) familly of DNA hydroxylases. The latter consists of 3 members TET1, 2 and 3. All 3 members catalyze the conversion of <strong>5-methylcytosine</strong> (5-mC) into <strong>5-hydroxymethylcytosine</strong> (5-hmC), and further into <strong>5-formylcytosine</strong> (5-fC) and <strong>5-carboxycytosine</strong> (5-caC). 5-fC and 5-caC can be converted to unmodified cytosine by <strong>Thymine DNA Glycosylase</strong> (TDG). It is not yet clear if 5-hmC, 5-fC and 5-caC have specific functions or are simply intermediates in the demethylation of 5-mC.</span></p>
<p><span style="font-weight: 400;">DNA methylation is generally considered as a repressive mark and is usually associated with gene silencing. It is essential that the balance between DNA methylation and demethylation is precisely maintained. Dysregulation of DNA methylation may lead to many different human diseases and is often observed in cancer cells.</span></p>
<p><span style="font-weight: 400;">Diagenode offers highly validated antibodies against different proteins involved in DNA modifications as well as against the modified bases allowing the study of all steps and intermediates in the DNA methylation/demethylation pathway:</span></p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/dna-methylation.jpg" height="599" width="816" /></p>
<p><strong>Diagenode exclusively sources the original 5-methylcytosine monoclonal antibody (clone 33D3).</strong></p>
<p>Check out the list below to see all proposed antibodies for DNA modifications.</p>
<p>Diagenode&rsquo;s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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			'description' => '<p><span style="font-weight: 400;">All Diagenode&rsquo;s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode&rsquo;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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			'name' => 'Developmental mRNA mC landscape and regulatory innovations of massivemC modification of maternal mRNAs in animals.',
			'authors' => 'Liu  J. et al.',
			'description' => '<p>mC is one of the longest-known RNA modifications, however, its developmental dynamics, functions, and evolution in mRNAs remain largely unknown. Here, we generate quantitative mRNA mC maps at different stages of development in 6 vertebrate and invertebrate species and find convergent and unexpected massive methylation of maternal mRNAs mediated by NSUN2 and NSUN6. Using Drosophila as a model, we reveal that embryos lacking maternal mRNA mC undergo cell cycle delays and fail to timely initiate maternal-to-zygotic transition, implying the functional importance of maternal mRNA mC. From invertebrates to the lineage leading to humans, two waves of mC regulatory innovations are observed: higher animals gain cis-directed NSUN2-mediated mC sites at the 5' end of the mRNAs, accompanied by the emergence of more structured 5'UTR regions; humans gain thousands of trans-directed NSUN6-mediated mC sites enriched in genes regulating the mitotic cell cycle. Collectively, our studies highlight the existence and regulatory innovations of a mechanism of early embryonic development and provide key resources for elucidating the role of mRNA mC in biology and disease.</p>',
			'date' => '2022-05-01',
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			'description' => '<p>Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.</p>',
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<p style="text-align: center;"><strong>Make your Bisulfite conversion now in only 60 minutes !</strong></p>
<p>Diagenode's Premium Bisulfite Kit rapidly converts DNA through bisulfite treatment. Our conversion reagent is added directly to DNA, requires no intermediate steps, and results in high yields of DNA ready for downstream analysis methods including PCR and Next-Generation Sequencing.</p>',
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	'description' => '<p>Hydroxymethylcytosine, well described in DNA, occurs also in RNA. Here, we show that hydroxymethylcytosine preferentially marks polyadenylated RNAs and is deposited by Tet in Drosophila. We map the transcriptome-wide hydroxymethylation landscape, revealing hydroxymethylcytosine in the transcripts of many genes, notably in coding sequences, and identify consensus sites for hydroxymethylation. We found that RNA hydroxymethylation can favor mRNA translation. Tet and hydroxymethylated RNA are found to be most abundant in the Drosophila brain, and Tet-deficient fruitflies suffer impaired brain development, accompanied by decreased RNA hydroxymethylation. This study highlights the distribution, localization, and function of cytosine hydroxymethylation and identifies central roles for this modification in Drosophila.</p>',
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include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118