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H2Bpan polyclonal antibody (sample size)

CUT&Tag grade Abs

ChIP-seq grade Abs

ChIP grade Abs

Negative IgG control

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Catalog Number

Format

Price

C15410157-10

10 µg

$115.00
Bulk order

Other format

C15410157

50 μg/18 μl

Polyclonal antibody raised in rabbit against histone H2B using a KLH-conjugated synthetic peptide containing an unmodified sequence from the C-terminal part of the protein.

Lot	A2622P
Concentration	2.8 µg/µl
Species reactivity	Human
Type	Polyclonal
Purity	Peptide affinity purified
Host	Rabbit
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.

Applications	Suggested dilution	References
ChIP ^*	0.5 - 1 µg/IP	Fig 1
ELISA	1:1,000 - 1/5,000	Fig 2
WB	1:2,000 - 1/10,000	Fig 3
IF	1:1,000	Fig 3

^* Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.

Validation data

Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B
ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.

Figure 2 Determination of the titer
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000.

Figure 3 Western blot analysis using the Diagenode antibody directed against H2B
Western blot was performed on whole cell extracts from HeLa cells (40 µg, lane 1) and on 1 µg of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B
HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) and with DAPI. Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 1% BSA. The cells were immunofluorescently labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
Target Description
Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histones play a central role in the regulation of transcription, DNA repair, DNA replication and chromosomal stability. These different functions are established via a complex set of post-translational modifications which either directly or indirectly alter chromatin structure and DNA accessibility to facilitate transcriptional activation or repression or other nuclear processes.

出版物

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HIRA supports hepatitis B virus minichromosome establishment andtranscriptional activity in infected hepatocytes.
Locatelli M. et al.
BACKGROUND \& AIMS: Upon Hepatitis B virus (HBV) infection, partially double stranded viral DNA converts into a covalently-closed-circular chromatinized episomal structure (cccDNA). This form represents the long-lived genomic reservoir responsible for viral persistence in the infected liver. While the involvemen...

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<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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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<td>Fig 1</td>
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<td>ELISA</td>
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<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-elisa.jpg" alt="H2Bpan Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-wb.jpg" alt="H2Bpan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-IF.jpg" alt="H2Bpan Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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			'description' => '<p><span style="font-weight: 400;">All Diagenode&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>
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			'meta_title' => 'Diagenode's selection of Antibodies is exclusively dedicated for Epigenetic Research | Diagenode',
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			'description' => '<h1><strong>Validated epigenetics antibodies</strong>&nbsp;&ndash; care for a sample?<br />&nbsp;</h1>
<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> -&nbsp;Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
<li><strong>Strict quality standards</strong>&nbsp;with rigorous QC and validation</li>
<li><strong>Classified</strong>&nbsp;based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. &nbsp;Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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			'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome.&nbsp; They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include&nbsp; methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation.&nbsp; The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>&rdquo;, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our&nbsp; antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode&rsquo;s collection includes antibodies recognizing:</p>
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<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
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<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode&rsquo;s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Expert technical support</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Sample sizes available</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 100% satisfaction guarantee</span></li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>).&nbsp;</p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" />&nbsp;</div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-chip.jpg" alt="H2Bpan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-elisa.jpg" alt="H2Bpan Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-wb.jpg" alt="H2Bpan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<div class="small-12 columns">
<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-elisa.jpg" alt="H2Bpan Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-wb.jpg" alt="H2Bpan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-IF.jpg" alt="H2Bpan Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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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	'meta_description' => 'Diagenode offers a wide range of antibodies and technical support for ChIP-qPCR applications',
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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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	'type' => 'Poster',
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	'modified' => '2024-01-16 16:48:37',
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	'name' => 'HIRA supports hepatitis B virus minichromosome establishment andtranscriptional activity in infected hepatocytes.',
	'authors' => 'Locatelli  M. et al.',
	'description' => '<p>BACKGROUND \&amp; AIMS: Upon Hepatitis B virus (HBV) infection, partially double stranded viral DNA converts into a covalently-closed-circular chromatinized episomal structure (cccDNA). This form represents the long-lived genomic reservoir responsible for viral persistence in the infected liver. While the involvement of host cell DNA damage response in cccDNA formation has been established, this work aims at investigating the yet to be identified histone dynamics on cccDNA during early phases of infection in human hepatocytes. METHODS: Detailed studies of host chromatin-associated factors were performed in cell culture models of natural infection, i.e. HepG2 cells and primary human hepatocytes infected with HBV, by cccDNA-specific chromatin immunoprecipitation and loss of function experiments during early kinetics of viral minichromosome establishment and onset of viral transcription. RESULTS: Our results show that cccDNA formation requires the deposition of the histone variant H3.3 via the histone regulator A (HIRA)-dependent pathway. This occurs simultaneously with repair of the cccDNA precursor and independently from de novo viral protein expression. Moreover, H3.3 in its S31 phosphorylated form appears to be the preferential H3 variant found on transcriptionally active cccDNA in infected cultured cells and human livers. HIRA depletion after cccDNA pool establishment demonstrated that HIRA recruitment is required for viral transcription and RNA production. CONCLUSIONS: Altogether, we show a crucial role for HIRA in the interplay between HBV genome and host cellular machinery to ensure the formation and active transcription of the viral minichromosome in infected hepatocytes.</p>',
	'date' => '2022-05-01',
	'pmid' => 'https://doi.org/10.1016%2Fj.jcmgh.2022.05.007',
	'doi' => '10.1016/j.jcmgh.2022.05.007',
	'modified' => '2022-08-11 14:38:05',
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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

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<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
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<td>ChIP <sup>*</sup></td>
<td>0.5 - 1 &micro;g/IP</td>
<td>Fig 1</td>
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<tr>
<td>ELISA</td>
<td>1:1,000 - 1/5,000</td>
<td>Fig 2</td>
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<td>WB</td>
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<td>Fig 3</td>
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<tr>
<td>IF</td>
<td>1:1,000</td>
<td>Fig 3</td>
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<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-elisa.jpg" alt="H2Bpan Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-wb.jpg" alt="H2Bpan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<div class="row">
<div class="small-12 columns">
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
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<li><strong>Focused</strong> -&nbsp;Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
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			'name' => 'Histone antibodies',
			'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome.&nbsp; They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include&nbsp; methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation.&nbsp; The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>&rdquo;, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our&nbsp; antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode&rsquo;s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode&rsquo;s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Expert technical support</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Sample sizes available</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 100% satisfaction guarantee</span></li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>).&nbsp;</p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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			'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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			'description' => '<p>BACKGROUND \&amp; AIMS: Upon Hepatitis B virus (HBV) infection, partially double stranded viral DNA converts into a covalently-closed-circular chromatinized episomal structure (cccDNA). This form represents the long-lived genomic reservoir responsible for viral persistence in the infected liver. While the involvement of host cell DNA damage response in cccDNA formation has been established, this work aims at investigating the yet to be identified histone dynamics on cccDNA during early phases of infection in human hepatocytes. METHODS: Detailed studies of host chromatin-associated factors were performed in cell culture models of natural infection, i.e. HepG2 cells and primary human hepatocytes infected with HBV, by cccDNA-specific chromatin immunoprecipitation and loss of function experiments during early kinetics of viral minichromosome establishment and onset of viral transcription. RESULTS: Our results show that cccDNA formation requires the deposition of the histone variant H3.3 via the histone regulator A (HIRA)-dependent pathway. This occurs simultaneously with repair of the cccDNA precursor and independently from de novo viral protein expression. Moreover, H3.3 in its S31 phosphorylated form appears to be the preferential H3 variant found on transcriptionally active cccDNA in infected cultured cells and human livers. HIRA depletion after cccDNA pool establishment demonstrated that HIRA recruitment is required for viral transcription and RNA production. CONCLUSIONS: Altogether, we show a crucial role for HIRA in the interplay between HBV genome and host cellular machinery to ensure the formation and active transcription of the viral minichromosome in infected hepatocytes.</p>',
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		'name' => 'H2Bpan Antibody',
		'description' => '<p>Polyclonal antibody raised in rabbit against <strong>histone H2B</strong> using a KLH-conjugated synthetic peptide containing an unmodified sequence from the C-terminal part of the protein.</p>',
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<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-chip.jpg" alt="H2Bpan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-7 columns">
<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-elisa.jpg" alt="H2Bpan Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-7 columns">
<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-wb.jpg" alt="H2Bpan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-IF.jpg" alt="H2Bpan Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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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	'name' => 'Antibodies you can trust',
	'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' => 'HIRA supports hepatitis B virus minichromosome establishment andtranscriptional activity in infected hepatocytes.',
	'authors' => 'Locatelli  M. et al.',
	'description' => '<p>BACKGROUND \&amp; AIMS: Upon Hepatitis B virus (HBV) infection, partially double stranded viral DNA converts into a covalently-closed-circular chromatinized episomal structure (cccDNA). This form represents the long-lived genomic reservoir responsible for viral persistence in the infected liver. While the involvement of host cell DNA damage response in cccDNA formation has been established, this work aims at investigating the yet to be identified histone dynamics on cccDNA during early phases of infection in human hepatocytes. METHODS: Detailed studies of host chromatin-associated factors were performed in cell culture models of natural infection, i.e. HepG2 cells and primary human hepatocytes infected with HBV, by cccDNA-specific chromatin immunoprecipitation and loss of function experiments during early kinetics of viral minichromosome establishment and onset of viral transcription. RESULTS: Our results show that cccDNA formation requires the deposition of the histone variant H3.3 via the histone regulator A (HIRA)-dependent pathway. This occurs simultaneously with repair of the cccDNA precursor and independently from de novo viral protein expression. Moreover, H3.3 in its S31 phosphorylated form appears to be the preferential H3 variant found on transcriptionally active cccDNA in infected cultured cells and human livers. HIRA depletion after cccDNA pool establishment demonstrated that HIRA recruitment is required for viral transcription and RNA production. CONCLUSIONS: Altogether, we show a crucial role for HIRA in the interplay between HBV genome and host cellular machinery to ensure the formation and active transcription of the viral minichromosome in infected hepatocytes.</p>',
	'date' => '2022-05-01',
	'pmid' => 'https://doi.org/10.1016%2Fj.jcmgh.2022.05.007',
	'doi' => '10.1016/j.jcmgh.2022.05.007',
	'modified' => '2022-08-11 14:38:05',
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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
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<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
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<tr>
<td>ChIP <sup>*</sup></td>
<td>0.5 - 1 &micro;g/IP</td>
<td>Fig 1</td>
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<tr>
<td>ELISA</td>
<td>1:1,000 - 1/5,000</td>
<td>Fig 2</td>
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<td>WB</td>
<td>1:2,000 - 1/10,000</td>
<td>Fig 3</td>
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<tr>
<td>IF</td>
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<td>Fig 3</td>
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<p><small><br /><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>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-chip.jpg" alt="H2Bpan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-wb.jpg" alt="H2Bpan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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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		'slug' => 'h2b-pan-polyclonal-antibody-classic-10-ug',
		'meta_title' => 'H2Bpan polyclonal antibody (sample size)',
		'meta_keywords' => '',
		'meta_description' => 'H2Bpan (Histone H2B) Polyclonal Antibody validated  in ChIP-qPCR, ELISA and WB. Batch-specific data available on the website.',
		'modified' => '2024-01-16 16:50:59',
		'created' => '2020-02-24 10:46:18',
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	),
	'Antibody' => array(
		'host' => '*****',
		'id' => '169',
		'name' => 'H2Bpan polyclonal antibody',
		'description' => 'Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histones play a central role in the regulation of transcription, DNA repair, DNA replication and chromosomal stability. These different functions are established via a complex set of post-translational modifications which either directly or indirectly alter chromatin structure and DNA accessibility to facilitate transcriptional activation or repression or other nuclear processes.',
		'clonality' => '',
		'isotype' => '',
		'lot' => 'A2622P',
		'concentration' => '2.8 &micro;g/&micro;l',
		'reactivity' => 'Human',
		'type' => 'Polyclonal',
		'purity' => 'Peptide affinity purified',
		'classification' => '',
		'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP <sup>*</sup></td>
<td>0.5 - 1 &micro;g/IP</td>
<td>Fig 1</td>
</tr>
<tr>
<td>ELISA</td>
<td>1:1,000 - 1/5,000</td>
<td>Fig 2</td>
</tr>
<tr>
<td>WB</td>
<td>1:2,000 - 1/10,000</td>
<td>Fig 3</td>
</tr>
<tr>
<td>IF</td>
<td>1:1,000</td>
<td>Fig 3</td>
</tr>
</tbody>
</table>
<p><small><br /><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>',
		'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' => '',
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		'modified' => '2018-07-03 17:18:13',
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			'name' => 'C15410157',
			'product_id' => '2255',
			'modified' => '2020-02-24 10:46:38',
			'created' => '2020-02-24 10:46:38'
		),
		'Master' => array(
			'id' => '2255',
			'antibody_id' => '169',
			'name' => 'H2Bpan Antibody',
			'description' => '<p>Polyclonal antibody raised in rabbit against <strong>histone H2B</strong> using a KLH-conjugated synthetic peptide containing an unmodified sequence from the C-terminal part of the protein.</p>',
			'label1' => 'Validation Data',
			'info1' => '<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-chip.jpg" alt="H2Bpan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-elisa.jpg" alt="H2Bpan Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-7 columns">
<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
</div>
</div>
<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-wb.jpg" alt="H2Bpan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-9 columns">
<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-IF.jpg" alt="H2Bpan Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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>',
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			'meta_description' => 'H2Bpan (Histone H2B) Polyclonal Antibody validated  in ChIP-qPCR, ELISA and WB. Batch-specific data available on the website.',
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			'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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			'meta_description' => 'Diagenode offers a wide range of antibodies and technical support for ChIP-qPCR applications',
			'meta_title' => 'ChIP Quantitative PCR  Antibodies (ChIP-qPCR) | Diagenode',
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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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			'meta_description' => 'Diagenode Offers Strict quality standards with Rigorous QC and validated Antibodies. Classified based on level of validation for flexibility of Application. Comprehensive selection of histone and non-histone Antibodies',
			'meta_title' => 'Diagenode's selection of Antibodies is exclusively dedicated for Epigenetic Research | Diagenode',
			'modified' => '2019-07-03 10:55:44',
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			'name' => 'Sample size antibodies',
			'description' => '<h1><strong>Validated epigenetics antibodies</strong>&nbsp;&ndash; care for a sample?<br />&nbsp;</h1>
<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
<ul>
<li><strong>Focused</strong> -&nbsp;Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
<li><strong>Strict quality standards</strong>&nbsp;with rigorous QC and validation</li>
<li><strong>Classified</strong>&nbsp;based on level of validation for flexibility of application</li>
</ul>
<p>Existing sample sizes are listed below. We will soon expand our collection. &nbsp;Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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			'meta_description' => 'Diagenode offers sample volume on selected antibodies for researchers to test, validate and provide confidence and flexibility in choosing from our wide range of antibodies ',
			'meta_title' => 'Sample-size Antibodies | Diagenode',
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			'name' => 'Histone antibodies',
			'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome.&nbsp; They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include&nbsp; methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation.&nbsp; The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>&rdquo;, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our&nbsp; antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode&rsquo;s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode&rsquo;s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Expert technical support</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Sample sizes available</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 100% satisfaction guarantee</span></li>
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			'meta_description' => 'Polyclonal and Monoclonal Antibodies against Histones and their modifications validated for many applications, including Chromatin Immunoprecipitation (ChIP) and ChIP-Sequencing (ChIP-seq)',
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			'name' => 'ChIP-grade antibodies',
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
</div>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>).&nbsp;</p>
<div class="row">
<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" />&nbsp;</div>
<div class="small-12 medium-6 large-6 columns">
<p></p>
<p></p>
<p></p>
</div>
</div>
<p></p>
<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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			'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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			'description' => '<p>BACKGROUND \&amp; AIMS: Upon Hepatitis B virus (HBV) infection, partially double stranded viral DNA converts into a covalently-closed-circular chromatinized episomal structure (cccDNA). This form represents the long-lived genomic reservoir responsible for viral persistence in the infected liver. While the involvement of host cell DNA damage response in cccDNA formation has been established, this work aims at investigating the yet to be identified histone dynamics on cccDNA during early phases of infection in human hepatocytes. METHODS: Detailed studies of host chromatin-associated factors were performed in cell culture models of natural infection, i.e. HepG2 cells and primary human hepatocytes infected with HBV, by cccDNA-specific chromatin immunoprecipitation and loss of function experiments during early kinetics of viral minichromosome establishment and onset of viral transcription. RESULTS: Our results show that cccDNA formation requires the deposition of the histone variant H3.3 via the histone regulator A (HIRA)-dependent pathway. This occurs simultaneously with repair of the cccDNA precursor and independently from de novo viral protein expression. Moreover, H3.3 in its S31 phosphorylated form appears to be the preferential H3 variant found on transcriptionally active cccDNA in infected cultured cells and human livers. HIRA depletion after cccDNA pool establishment demonstrated that HIRA recruitment is required for viral transcription and RNA production. CONCLUSIONS: Altogether, we show a crucial role for HIRA in the interplay between HBV genome and host cellular machinery to ensure the formation and active transcription of the viral minichromosome in infected hepatocytes.</p>',
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<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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	'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' => 'SDS C15410157 H2Bpan Antibody DE de',
	'language' => 'de',
	'url' => 'files/SDS/H2Bpan/SDS-C15410157-H2Bpan_Antibody-DE-de-GHS_2_0b.pdf',
	'countries' => 'DE',
	'modified' => '2024-01-16 16:48:37',
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	'name' => 'HIRA supports hepatitis B virus minichromosome establishment andtranscriptional activity in infected hepatocytes.',
	'authors' => 'Locatelli  M. et al.',
	'description' => '<p>BACKGROUND \&amp; AIMS: Upon Hepatitis B virus (HBV) infection, partially double stranded viral DNA converts into a covalently-closed-circular chromatinized episomal structure (cccDNA). This form represents the long-lived genomic reservoir responsible for viral persistence in the infected liver. While the involvement of host cell DNA damage response in cccDNA formation has been established, this work aims at investigating the yet to be identified histone dynamics on cccDNA during early phases of infection in human hepatocytes. METHODS: Detailed studies of host chromatin-associated factors were performed in cell culture models of natural infection, i.e. HepG2 cells and primary human hepatocytes infected with HBV, by cccDNA-specific chromatin immunoprecipitation and loss of function experiments during early kinetics of viral minichromosome establishment and onset of viral transcription. RESULTS: Our results show that cccDNA formation requires the deposition of the histone variant H3.3 via the histone regulator A (HIRA)-dependent pathway. This occurs simultaneously with repair of the cccDNA precursor and independently from de novo viral protein expression. Moreover, H3.3 in its S31 phosphorylated form appears to be the preferential H3 variant found on transcriptionally active cccDNA in infected cultured cells and human livers. HIRA depletion after cccDNA pool establishment demonstrated that HIRA recruitment is required for viral transcription and RNA production. CONCLUSIONS: Altogether, we show a crucial role for HIRA in the interplay between HBV genome and host cellular machinery to ensure the formation and active transcription of the viral minichromosome in infected hepatocytes.</p>',
	'date' => '2022-05-01',
	'pmid' => 'https://doi.org/10.1016%2Fj.jcmgh.2022.05.007',
	'doi' => '10.1016/j.jcmgh.2022.05.007',
	'modified' => '2022-08-11 14:38:05',
	'created' => '2022-08-11 12:14:50',
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$externalLink = ' <a href="https://doi.org/10.1016%2Fj.jcmgh.2022.05.007" target="_blank"><i class="fa fa-external-link"></i></a>'
include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118

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<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-chip.jpg" alt="H2Bpan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<div class="row">
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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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<th>References</th>
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<td>Fig 1</td>
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<tr>
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<td>Fig 2</td>
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<tr>
<td>WB</td>
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<td>Fig 3</td>
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<td>IF</td>
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<td>Fig 3</td>
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<div class="small-5 columns">
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<div class="small-7 columns">
<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-elisa.jpg" alt="H2Bpan Antibody ELISA validation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-7 columns">
<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
</div>
</div>
<div class="row">
<div class="small-3 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-wb.jpg" alt="H2Bpan Antibody validated in Western Blot" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-9 columns">
<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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</div>
<div class="row">
<div class="small-12 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-IF.jpg" alt="H2Bpan Antibody validated in Immunofluorescence" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
</div>
<div class="row">
<div class="small-12 columns">
<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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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			'meta_description' => 'H2Bpan (Histone H2B) Polyclonal Antibody validated  in ChIP-qPCR, ELISA and WB. Batch-specific data available on the website.',
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			'description' => '<p><strong>Western blot</strong> : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.</p>
<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
<p><em></em>Check our selection of antibodies validated in Western blot.</p>',
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			'description' => '<p><span style="font-weight: 400;">All Diagenode&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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			'meta_description' => 'Diagenode Offers Strict quality standards with Rigorous QC and validated Antibodies. Classified based on level of validation for flexibility of Application. Comprehensive selection of histone and non-histone Antibodies',
			'meta_title' => 'Diagenode's selection of Antibodies is exclusively dedicated for Epigenetic Research | Diagenode',
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			'description' => '<h1><strong>Validated epigenetics antibodies</strong>&nbsp;&ndash; care for a sample?<br />&nbsp;</h1>
<p>Diagenode has partnered with leading epigenetics experts and numerous epigenetics consortiums to bring to you a validated and comprehensive collection of epigenetic antibodies. As an expert in epigenetics, we are committed to offering highly-specific antibodies validated for ChIP/ChIP-seq and many other applications. All batch-specific validation data is available on our website.<br /><a href="../categories/antibodies">Read about our expertise in antibody production</a>.</p>
<ul>
<li><strong>Focused</strong> -&nbsp;Diagenode's selection of antibodies is exclusively dedicated for epigenetic research. <a title="See the full collection." href="../categories/all-antibodies">See the full collection.</a></li>
<li><strong>Strict quality standards</strong>&nbsp;with rigorous QC and validation</li>
<li><strong>Classified</strong>&nbsp;based on level of validation for flexibility of application</li>
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<p>Existing sample sizes are listed below. We will soon expand our collection. &nbsp;Are you looking for a sample size of another antibody? Just <a href="mailto:agnieszka.zelisko@diagenode.com?Subject=Sample%20Size%20Request" target="_top">Contact us</a>.</p>',
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			'name' => 'Histone antibodies',
			'description' => '<p>Histones are the main protein components of chromatin involved in the compaction of DNA into nucleosomes, the basic units of chromatin. A <strong>nucleosome</strong> consists of one pair of each of the core histones (<strong>H2A</strong>, <strong>H2B</strong>, <strong>H3</strong> and <strong>H4</strong>) forming an octameric structure wrapped by 146 base pairs of DNA. The different nucleosomes are linked by the linker histone<strong> H1, </strong>allowing for further condensation of chromatin.</p>
<p>The core histones have a globular structure with large unstructured N-terminal tails protruding from the nucleosome.&nbsp; They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include&nbsp; methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation.&nbsp; The most well characterized PTMs are <strong>methylation,</strong> <strong>acetylation and phosphorylation</strong>. Histone methylation occurs mainly on lysine (K) residues, which can be mono-, di- or tri-methylated, and on arginines (R), which can be mono-methylated and symmetrically or asymmetrically di-methylated. Histone acetylation occurs on lysines and histone phosphorylation mainly on serines (S), threonines (T) and tyrosines (Y).</p>
<p>The PTMs of the different residues are involved in numerous processes such as DNA repair, DNA replication and chromosome condensation. They influence the chromatin organization and can be positively or negatively associated with gene expression. Trimethylation of H3K4, H3K36 and H3K79, and lysine acetylation generally result in an open chromatin configuration (figure below) and are therefore associated with <strong>euchromatin</strong> and gene activation. Trimethylation of H3K9, K3K27 and H4K20, on the other hand, is enriched in <strong>heterochromatin </strong>and associated with gene silencing. The combination of different histone modifications is called the "<strong>histone code</strong>&rdquo;, analogous to the genetic code.</p>
<p><img src="https://www.diagenode.com/img/categories/antibodies/histone-marks-illustration.png" /></p>
<p>Diagenode is proud to offer a large range of antibodies against histones and histone modifications. Our&nbsp; antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode&rsquo;s collection includes antibodies recognizing:</p>
<ul>
<li><strong>Histone H1 variants</strong></li>
<li><strong>Histone H2A, H2A variants and histone H2A</strong> <strong>modifications</strong> (serine phosphorylation, lysine acetylation, lysine ubiquitinylation)</li>
<li><strong>Histone H2B and H2B</strong> <strong>modifications </strong>(serine phosphorylation, lysine acetylation)</li>
<li><strong>Histone H3 and H3 modifications </strong>(lysine methylation (mono-, di- and tri-methylated), lysine acetylation, serine phosphorylation, threonine phosphorylation, arginine methylation (mono-methylated, symmetrically and asymmetrically di-methylated))</li>
<li><strong>Histone H4 and H4 modifications (</strong>lysine methylation (mono-, di- and tri-methylated), lysine acetylation, arginine methylation (mono-methylated and symmetrically di-methylated), serine phosphorylation )</li>
</ul>
<p><span style="font-weight: 400;"><strong>HDAC's HAT's, HMT's and other</strong> <strong>enzymes</strong> which modify histones can be found in the category <a href="../categories/chromatin-modifying-proteins-histone-transferase">Histone modifying enzymes</a><br /></span></p>
<p><span style="font-weight: 400;"> Diagenode&rsquo;s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Expert technical support</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Sample sizes available</span></li>
<li><span style="font-weight: 400;"> &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; 100% satisfaction guarantee</span></li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>).&nbsp;</p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" />&nbsp;</div>
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<p></p>
<p></p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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			'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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			'authors' => 'Locatelli  M. et al.',
			'description' => '<p>BACKGROUND \&amp; AIMS: Upon Hepatitis B virus (HBV) infection, partially double stranded viral DNA converts into a covalently-closed-circular chromatinized episomal structure (cccDNA). This form represents the long-lived genomic reservoir responsible for viral persistence in the infected liver. While the involvement of host cell DNA damage response in cccDNA formation has been established, this work aims at investigating the yet to be identified histone dynamics on cccDNA during early phases of infection in human hepatocytes. METHODS: Detailed studies of host chromatin-associated factors were performed in cell culture models of natural infection, i.e. HepG2 cells and primary human hepatocytes infected with HBV, by cccDNA-specific chromatin immunoprecipitation and loss of function experiments during early kinetics of viral minichromosome establishment and onset of viral transcription. RESULTS: Our results show that cccDNA formation requires the deposition of the histone variant H3.3 via the histone regulator A (HIRA)-dependent pathway. This occurs simultaneously with repair of the cccDNA precursor and independently from de novo viral protein expression. Moreover, H3.3 in its S31 phosphorylated form appears to be the preferential H3 variant found on transcriptionally active cccDNA in infected cultured cells and human livers. HIRA depletion after cccDNA pool establishment demonstrated that HIRA recruitment is required for viral transcription and RNA production. CONCLUSIONS: Altogether, we show a crucial role for HIRA in the interplay between HBV genome and host cellular machinery to ensure the formation and active transcription of the viral minichromosome in infected hepatocytes.</p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410157-chip.jpg" alt="H2Bpan Antibody ChIP Grade" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><small><strong>Figure 1 ChIP results obtained with the Diagenode antibody directed against H2B</strong><br /> ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2B (cat. No. C15410157) and optimized PCR primer sets for qPCR. ChIP was performed with the Auto Histone ChIP-seq kit (Cat. No. C01010022) on sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 0.5, 1, 2 and 5 &micro;g per ChIP experiment was analysed. IgG (2 &micro;g/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.</small></p>
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<p><small><strong>Figure 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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<p><small><strong>Figure 4 Immunofluorescence using the Diagenode antibody directed against H2B</strong><br />HeLa cells were stained with the Diagenode antibody against H2B (Cat. No. C15410157) 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 labeled with the H2B antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. 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 2 Determination of the titer</strong><br /> To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against H2B (C15410157) in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:40,000. </small></p>
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<p><small><strong>Figure 3 Western blot analysis using the Diagenode antibody directed against H2B</strong><br /> Western blot was performed on whole cell extracts from HeLa cells (40 &micro;g, lane 1) and on 1 &micro;g of recombinant histone H2B (lane 2) using the Diagenode antibody against H2B (Cat. No. C15410157). The antibody was diluted 1:10,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.</small></p>
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	'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' => 'HIRA supports hepatitis B virus minichromosome establishment andtranscriptional activity in infected hepatocytes.',
	'authors' => 'Locatelli  M. et al.',
	'description' => '<p>BACKGROUND \&amp; AIMS: Upon Hepatitis B virus (HBV) infection, partially double stranded viral DNA converts into a covalently-closed-circular chromatinized episomal structure (cccDNA). This form represents the long-lived genomic reservoir responsible for viral persistence in the infected liver. While the involvement of host cell DNA damage response in cccDNA formation has been established, this work aims at investigating the yet to be identified histone dynamics on cccDNA during early phases of infection in human hepatocytes. METHODS: Detailed studies of host chromatin-associated factors were performed in cell culture models of natural infection, i.e. HepG2 cells and primary human hepatocytes infected with HBV, by cccDNA-specific chromatin immunoprecipitation and loss of function experiments during early kinetics of viral minichromosome establishment and onset of viral transcription. RESULTS: Our results show that cccDNA formation requires the deposition of the histone variant H3.3 via the histone regulator A (HIRA)-dependent pathway. This occurs simultaneously with repair of the cccDNA precursor and independently from de novo viral protein expression. Moreover, H3.3 in its S31 phosphorylated form appears to be the preferential H3 variant found on transcriptionally active cccDNA in infected cultured cells and human livers. HIRA depletion after cccDNA pool establishment demonstrated that HIRA recruitment is required for viral transcription and RNA production. CONCLUSIONS: Altogether, we show a crucial role for HIRA in the interplay between HBV genome and host cellular machinery to ensure the formation and active transcription of the viral minichromosome in infected hepatocytes.</p>',
	'date' => '2022-05-01',
	'pmid' => 'https://doi.org/10.1016%2Fj.jcmgh.2022.05.007',
	'doi' => '10.1016/j.jcmgh.2022.05.007',
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View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
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Antibodies you can trust POSTER Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...	Download

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