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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--ChIP.png" alt="H4K20me1 Antibody ChIP-seq" caption="false" width="288" height="219" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade " caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq " caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" caption="false" width="367" height="60" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" caption="false" width="288" height="226" /></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" caption="false" width="280" height="180" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" caption="false" width="367" height="139" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" caption="false" width="367" height="139" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<td>ChIP/ChIP-seq <sup>*</sup></td>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--ChIP.png" alt="H4K20me1 Antibody ChIP-seq" caption="false" width="288" height="219" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" caption="false" width="367" height="139" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" caption="false" width="367" height="139" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<td>Fig 1, 2</td>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<div class="row">
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>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. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. 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>”, 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 antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’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’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
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<p>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> - 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> with rigorous QC and validation</li>
<li><strong>Classified</strong> 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. 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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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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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>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'authors' => 'Ugai, Keita and Matsuda, Shuichi and Mikami, Hideki and Shimada, Ayako andMisawa, Tomoko and Nakamura, Hiroyuki and Tatsumi, Koichiro and Hatano,Masahiko and Murayama, Toshihiko and Kasuya, Yoshitoshi',
'description' => '<p>Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease of unknown etiopathogenesis. The activation of extracellular matrix (ECM)-producing myofibroblasts plays a key role in fibrotic tissue remodeling. The dedifferentiation of myofibroblasts has attracted considerable attention as a promising target for the development of effective therapeutic interventions against IPF. Here, we screened a small library of epigenetics-related inhibitors using dedifferentiation assay of lung myofibroblasts prepared from a patient at the terminal stages of IPF and chose UNC0379. The inhibition of SET8, a histone H4 lysine 20 (H4K20) monomethyltransferase, by UNC0379 markedly suppressed the expression of α-smooth muscle actin (SMA) and ED-A-fibronectin in myofibroblasts. In IPF myofibroblasts, SET8 expression and H4K20 monomethylation (H4K20me1) levels, which were significantly higher than those in normal human lung fibroblasts, were reduced upon treatment with UNC0379. Hence, the changes in the expression of the two fibrotic markers clearly correlated with those in SET8 expression and H4K20me1 level. Furthermore, in a mouse model of bleomycin (BLM)-induced lung fibrosis, the intratracheal administration of UNC0379 at an early fibrotic stage markedly ameliorated the histopathological changes associated with collagen deposition in the lungs. However, treatment with UNC0379 did not significantly affect the number of proinflammatory cells or cytokine production in the bronchoalveolar lavage fluids from mice treated with BLM. In the BLM-injured lung, SET8 was predominantly localized to the nuclei of α-SMA-positive cells, which colocalized with H4K20me1. Taken together, our results indicate that the inhibition of {SET}8 resulting in myofibroblast dedifferentiation may partly mitigate lung fibrosis without affecting the inflammatory responses.</p>',
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'authors' => 'Piazza R, Magistroni V, Redaelli S, Mauri M, Massimino L, Sessa A, Peronaci M, Lalowski M, Soliymani R, Mezzatesta C, Pirola A, Banfi F, Rubio A, Rea D, Stagno F, Usala E, Martino B, Campiotti L, Merli M, Passamonti F, Onida F, Morotti A, Pavesi F, Bregni',
'description' => '<p>SETBP1 variants occur as somatic mutations in several hematological malignancies such as atypical chronic myeloid leukemia and as de novo germline mutations in the Schinzel-Giedion syndrome. Here we show that SETBP1 binds to gDNA in AT-rich promoter regions, causing activation of gene expression through recruitment of a HCF1/KMT2A/PHF8 epigenetic complex. Deletion of two AT-hooks abrogates the binding of SETBP1 to gDNA and impairs target gene upregulation. Genes controlled by SETBP1 such as MECOM are significantly upregulated in leukemias containing SETBP1 mutations. Gene ontology analysis of deregulated SETBP1 target genes indicates that they are also key controllers of visceral organ development and brain morphogenesis. In line with these findings, in utero brain electroporation of mutated SETBP1 causes impairment of mouse neurogenesis with a profound delay in neuronal migration. In summary, this work unveils a SETBP1 function that directly affects gene transcription and clarifies the mechanism operating in myeloid malignancies and in the Schinzel-Giedion syndrome caused by SETBP1 mutations.</p>',
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'description' => 'DNA double-strand breaks (DSBs) activate a signaling pathway known as the DNA damage response (DDR) which via protein–protein interactions and post-translational modifications recruit signaling proteins, such as 53BP1, to chromatin flanking the lesion. Depletion of the SET8 methyltransferase prevents accumulation of 53BP1 at DSBs; however, this phenotype has been attributed to the role of SET8 in generating H4K20 methylation across the genome, which is required for 53BP1 binding to chromatin, prior to DNA damage. Here, we report that SET8 acts directly at DSBs during the DNA damage response (DDR). SET8 accumulates at DSBs and is enzymatically active at DSBs. Depletion of SET8 just prior to the induction of DNA damage abrogates 53BP1's accumulation at DSBs, suggesting that SET8 acts during DDR. SET8's occupancy at DSBs is regulated by histone deacetylases (HDACs). Finally, SET8 is functionally required for efficient repair of DSBs specifically via the non-homologous end-joining pathway (NHEJ). Our findings reveal that SET8's active role during DDR at DSBs is required for 53BP1's accumulation.',
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'name' => 'H4K20me1 Antibody',
'description' => '<p><span>Monoclonal antibody raised in mouse against histone <strong>H4 containing the monomethylated lysine 20 (H4K20me1)</strong>, using a KLH-conjugated synthetic peptide. </span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--ChIP.png" alt="H4K20me1 Antibody ChIP Grade" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade " caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq " caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" caption="false" width="367" height="60" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" caption="false" width="288" height="226" /></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" caption="false" width="280" height="180" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" caption="false" width="367" height="139" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" caption="false" width="367" height="139" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" caption="false" width="280" height="180" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" caption="false" width="367" height="139" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" caption="false" width="367" height="139" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
</div>
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'application_table' => '<table>
<thead>
<tr>
<th>Applications</th>
<th>Suggested dilution</th>
<th>References</th>
</tr>
</thead>
<tbody>
<tr>
<td>ChIP/ChIP-seq <sup>*</sup></td>
<td>0,5 - 1 μg/ChIP</td>
<td>Fig 1, 2</td>
</tr>
<tr>
<td>Dot Blotting</td>
<td>1:20,000</td>
<td>Fig 3</td>
</tr>
<tr>
<td>Western Blotting</td>
<td>1:1,000</td>
<td>Fig 4</td>
</tr>
<tr>
<td>Immunofluorescence</td>
<td>1:200</td>
<td>Fig 5</td>
</tr>
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<p><small><sup>*</sup> Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 0.5-5 μg per IP.</small></p>',
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'description' => '<p><span>Monoclonal antibody raised in mouse against histone <strong>H4 containing the monomethylated lysine 20 (H4K20me1)</strong>, using a KLH-conjugated synthetic peptide. </span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--ChIP.png" alt="H4K20me1 Antibody ChIP Grade" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>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. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. 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>”, 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 antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’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>
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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’s highly validated antibodies:</span></p>
<ul>
<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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<p>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> - 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> with rigorous QC and validation</li>
<li><strong>Classified</strong> 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. 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><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
<ul>
<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
<li>Sample sizes available</li>
<li>100% satisfaction guarantee</li>
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<div class="small-12 columns"><center></center>
<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
</div>
</div>
<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p></p>
<p></p>
<p></p>
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<p></p>
<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'authors' => 'Ugai, Keita and Matsuda, Shuichi and Mikami, Hideki and Shimada, Ayako andMisawa, Tomoko and Nakamura, Hiroyuki and Tatsumi, Koichiro and Hatano,Masahiko and Murayama, Toshihiko and Kasuya, Yoshitoshi',
'description' => '<p>Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease of unknown etiopathogenesis. The activation of extracellular matrix (ECM)-producing myofibroblasts plays a key role in fibrotic tissue remodeling. The dedifferentiation of myofibroblasts has attracted considerable attention as a promising target for the development of effective therapeutic interventions against IPF. Here, we screened a small library of epigenetics-related inhibitors using dedifferentiation assay of lung myofibroblasts prepared from a patient at the terminal stages of IPF and chose UNC0379. The inhibition of SET8, a histone H4 lysine 20 (H4K20) monomethyltransferase, by UNC0379 markedly suppressed the expression of α-smooth muscle actin (SMA) and ED-A-fibronectin in myofibroblasts. In IPF myofibroblasts, SET8 expression and H4K20 monomethylation (H4K20me1) levels, which were significantly higher than those in normal human lung fibroblasts, were reduced upon treatment with UNC0379. Hence, the changes in the expression of the two fibrotic markers clearly correlated with those in SET8 expression and H4K20me1 level. Furthermore, in a mouse model of bleomycin (BLM)-induced lung fibrosis, the intratracheal administration of UNC0379 at an early fibrotic stage markedly ameliorated the histopathological changes associated with collagen deposition in the lungs. However, treatment with UNC0379 did not significantly affect the number of proinflammatory cells or cytokine production in the bronchoalveolar lavage fluids from mice treated with BLM. In the BLM-injured lung, SET8 was predominantly localized to the nuclei of α-SMA-positive cells, which colocalized with H4K20me1. Taken together, our results indicate that the inhibition of {SET}8 resulting in myofibroblast dedifferentiation may partly mitigate lung fibrosis without affecting the inflammatory responses.</p>',
'date' => '2020-08-05',
'pmid' => 'https://www.frontiersin.org/articles/10.3389/fmolb.2020.00192/full',
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'authors' => 'Piazza R, Magistroni V, Redaelli S, Mauri M, Massimino L, Sessa A, Peronaci M, Lalowski M, Soliymani R, Mezzatesta C, Pirola A, Banfi F, Rubio A, Rea D, Stagno F, Usala E, Martino B, Campiotti L, Merli M, Passamonti F, Onida F, Morotti A, Pavesi F, Bregni',
'description' => '<p>SETBP1 variants occur as somatic mutations in several hematological malignancies such as atypical chronic myeloid leukemia and as de novo germline mutations in the Schinzel-Giedion syndrome. Here we show that SETBP1 binds to gDNA in AT-rich promoter regions, causing activation of gene expression through recruitment of a HCF1/KMT2A/PHF8 epigenetic complex. Deletion of two AT-hooks abrogates the binding of SETBP1 to gDNA and impairs target gene upregulation. Genes controlled by SETBP1 such as MECOM are significantly upregulated in leukemias containing SETBP1 mutations. Gene ontology analysis of deregulated SETBP1 target genes indicates that they are also key controllers of visceral organ development and brain morphogenesis. In line with these findings, in utero brain electroporation of mutated SETBP1 causes impairment of mouse neurogenesis with a profound delay in neuronal migration. In summary, this work unveils a SETBP1 function that directly affects gene transcription and clarifies the mechanism operating in myeloid malignancies and in the Schinzel-Giedion syndrome caused by SETBP1 mutations.</p>',
'date' => '2018-06-06',
'pmid' => 'http://www.pubmed.gov/29875417',
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'name' => 'SET8 methyltransferase activity during the DNA double-strand break response is required for recruitment of 53BP1',
'authors' => 'Dulev, S., Tkach, J., Lin, S. and Batada, N. N.',
'description' => 'DNA double-strand breaks (DSBs) activate a signaling pathway known as the DNA damage response (DDR) which via protein–protein interactions and post-translational modifications recruit signaling proteins, such as 53BP1, to chromatin flanking the lesion. Depletion of the SET8 methyltransferase prevents accumulation of 53BP1 at DSBs; however, this phenotype has been attributed to the role of SET8 in generating H4K20 methylation across the genome, which is required for 53BP1 binding to chromatin, prior to DNA damage. Here, we report that SET8 acts directly at DSBs during the DNA damage response (DDR). SET8 accumulates at DSBs and is enzymatically active at DSBs. Depletion of SET8 just prior to the induction of DNA damage abrogates 53BP1's accumulation at DSBs, suggesting that SET8 acts during DDR. SET8's occupancy at DSBs is regulated by histone deacetylases (HDACs). Finally, SET8 is functionally required for efficient repair of DSBs specifically via the non-homologous end-joining pathway (NHEJ). Our findings reveal that SET8's active role during DDR at DSBs is required for 53BP1's accumulation.',
'date' => '2014-09-25',
'pmid' => 'http://onlinelibrary.wiley.com/doi/10.15252/embr.201439434/abstract',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" caption="false" width="367" height="139" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" caption="false" width="367" height="139" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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</div>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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'description' => 'DNA double-strand breaks (DSBs) activate a signaling pathway known as the DNA damage response (DDR) which via protein–protein interactions and post-translational modifications recruit signaling proteins, such as 53BP1, to chromatin flanking the lesion. Depletion of the SET8 methyltransferase prevents accumulation of 53BP1 at DSBs; however, this phenotype has been attributed to the role of SET8 in generating H4K20 methylation across the genome, which is required for 53BP1 binding to chromatin, prior to DNA damage. Here, we report that SET8 acts directly at DSBs during the DNA damage response (DDR). SET8 accumulates at DSBs and is enzymatically active at DSBs. Depletion of SET8 just prior to the induction of DNA damage abrogates 53BP1's accumulation at DSBs, suggesting that SET8 acts during DDR. SET8's occupancy at DSBs is regulated by histone deacetylases (HDACs). Finally, SET8 is functionally required for efficient repair of DSBs specifically via the non-homologous end-joining pathway (NHEJ). Our findings reveal that SET8's active role during DDR at DSBs is required for 53BP1's accumulation.',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" caption="false" width="367" height="139" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" caption="false" width="367" height="139" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<div class="small-5 columns">
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </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><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>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. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. 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>”, 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 antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’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’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
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<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
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<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
<li>Expert technical support</li>
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<p>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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<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'description' => '<p style="text-align: justify;"><span>Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of large sets of genome-wide sequencing data. ChIP sequencing (ChIP-seq) has now become the gold standard method for chromatin studies, given the accuracy and coverage scale of the approach over other methods. Successful ChIP-seq, however, requires a higher level of experimental accuracy and consistency in all steps of ChIP than ever before. Particularly crucial is the quality of ChIP antibodies. </span></p>',
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'name' => 'Inhibition of the SET8 Pathway Ameliorates Lung Fibrosis Even ThroughFibroblast Dedifferentiation',
'authors' => 'Ugai, Keita and Matsuda, Shuichi and Mikami, Hideki and Shimada, Ayako andMisawa, Tomoko and Nakamura, Hiroyuki and Tatsumi, Koichiro and Hatano,Masahiko and Murayama, Toshihiko and Kasuya, Yoshitoshi',
'description' => '<p>Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease of unknown etiopathogenesis. The activation of extracellular matrix (ECM)-producing myofibroblasts plays a key role in fibrotic tissue remodeling. The dedifferentiation of myofibroblasts has attracted considerable attention as a promising target for the development of effective therapeutic interventions against IPF. Here, we screened a small library of epigenetics-related inhibitors using dedifferentiation assay of lung myofibroblasts prepared from a patient at the terminal stages of IPF and chose UNC0379. The inhibition of SET8, a histone H4 lysine 20 (H4K20) monomethyltransferase, by UNC0379 markedly suppressed the expression of α-smooth muscle actin (SMA) and ED-A-fibronectin in myofibroblasts. In IPF myofibroblasts, SET8 expression and H4K20 monomethylation (H4K20me1) levels, which were significantly higher than those in normal human lung fibroblasts, were reduced upon treatment with UNC0379. Hence, the changes in the expression of the two fibrotic markers clearly correlated with those in SET8 expression and H4K20me1 level. Furthermore, in a mouse model of bleomycin (BLM)-induced lung fibrosis, the intratracheal administration of UNC0379 at an early fibrotic stage markedly ameliorated the histopathological changes associated with collagen deposition in the lungs. However, treatment with UNC0379 did not significantly affect the number of proinflammatory cells or cytokine production in the bronchoalveolar lavage fluids from mice treated with BLM. In the BLM-injured lung, SET8 was predominantly localized to the nuclei of α-SMA-positive cells, which colocalized with H4K20me1. Taken together, our results indicate that the inhibition of {SET}8 resulting in myofibroblast dedifferentiation may partly mitigate lung fibrosis without affecting the inflammatory responses.</p>',
'date' => '2020-08-05',
'pmid' => 'https://www.frontiersin.org/articles/10.3389/fmolb.2020.00192/full',
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'authors' => 'Piazza R, Magistroni V, Redaelli S, Mauri M, Massimino L, Sessa A, Peronaci M, Lalowski M, Soliymani R, Mezzatesta C, Pirola A, Banfi F, Rubio A, Rea D, Stagno F, Usala E, Martino B, Campiotti L, Merli M, Passamonti F, Onida F, Morotti A, Pavesi F, Bregni',
'description' => '<p>SETBP1 variants occur as somatic mutations in several hematological malignancies such as atypical chronic myeloid leukemia and as de novo germline mutations in the Schinzel-Giedion syndrome. Here we show that SETBP1 binds to gDNA in AT-rich promoter regions, causing activation of gene expression through recruitment of a HCF1/KMT2A/PHF8 epigenetic complex. Deletion of two AT-hooks abrogates the binding of SETBP1 to gDNA and impairs target gene upregulation. Genes controlled by SETBP1 such as MECOM are significantly upregulated in leukemias containing SETBP1 mutations. Gene ontology analysis of deregulated SETBP1 target genes indicates that they are also key controllers of visceral organ development and brain morphogenesis. In line with these findings, in utero brain electroporation of mutated SETBP1 causes impairment of mouse neurogenesis with a profound delay in neuronal migration. In summary, this work unveils a SETBP1 function that directly affects gene transcription and clarifies the mechanism operating in myeloid malignancies and in the Schinzel-Giedion syndrome caused by SETBP1 mutations.</p>',
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'pmid' => 'http://www.pubmed.gov/29875417',
'doi' => '10.1038/s41467-018-04462-8',
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'authors' => 'Dulev, S., Tkach, J., Lin, S. and Batada, N. N.',
'description' => 'DNA double-strand breaks (DSBs) activate a signaling pathway known as the DNA damage response (DDR) which via protein–protein interactions and post-translational modifications recruit signaling proteins, such as 53BP1, to chromatin flanking the lesion. Depletion of the SET8 methyltransferase prevents accumulation of 53BP1 at DSBs; however, this phenotype has been attributed to the role of SET8 in generating H4K20 methylation across the genome, which is required for 53BP1 binding to chromatin, prior to DNA damage. Here, we report that SET8 acts directly at DSBs during the DNA damage response (DDR). SET8 accumulates at DSBs and is enzymatically active at DSBs. Depletion of SET8 just prior to the induction of DNA damage abrogates 53BP1's accumulation at DSBs, suggesting that SET8 acts during DDR. SET8's occupancy at DSBs is regulated by histone deacetylases (HDACs). Finally, SET8 is functionally required for efficient repair of DSBs specifically via the non-homologous end-joining pathway (NHEJ). Our findings reveal that SET8's active role during DDR at DSBs is required for 53BP1's accumulation.',
'date' => '2014-09-25',
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'name' => 'H4K20me1 Antibody',
'description' => '<p><span>Monoclonal antibody raised in mouse against histone <strong>H4 containing the monomethylated lysine 20 (H4K20me1)</strong>, using a KLH-conjugated synthetic peptide. </span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--ChIP.png" alt="H4K20me1 Antibody ChIP Grade" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade " caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq " caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" caption="false" width="367" height="60" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" caption="false" width="367" height="60" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" caption="false" width="288" height="226" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" caption="false" width="280" height="180" /></p>
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<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" caption="false" width="367" height="139" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" caption="false" width="367" height="139" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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'description' => 'DNA double-strand breaks (DSBs) activate a signaling pathway known as the DNA damage response (DDR) which via protein–protein interactions and post-translational modifications recruit signaling proteins, such as 53BP1, to chromatin flanking the lesion. Depletion of the SET8 methyltransferase prevents accumulation of 53BP1 at DSBs; however, this phenotype has been attributed to the role of SET8 in generating H4K20 methylation across the genome, which is required for 53BP1 binding to chromatin, prior to DNA damage. Here, we report that SET8 acts directly at DSBs during the DNA damage response (DDR). SET8 accumulates at DSBs and is enzymatically active at DSBs. Depletion of SET8 just prior to the induction of DNA damage abrogates 53BP1's accumulation at DSBs, suggesting that SET8 acts during DDR. SET8's occupancy at DSBs is regulated by histone deacetylases (HDACs). Finally, SET8 is functionally required for efficient repair of DSBs specifically via the non-homologous end-joining pathway (NHEJ). Our findings reveal that SET8's active role during DDR at DSBs is required for 53BP1's accumulation.',
'date' => '2014-09-25',
'pmid' => 'http://onlinelibrary.wiley.com/doi/10.15252/embr.201439434/abstract',
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'modified' => '2015-07-24 15:39:03',
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p>Learn more about: <a href="https://www.diagenode.com/applications/western-blot">Loading control, MW marker visualization</a><em>. <br /></em></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200229-IF.jpg" alt="" height="245" width="256" /></p>
<p><sup><strong>Immunofluorescence using the Diagenode monoclonal antibody directed against CRISPR/Cas9</strong></sup></p>
<p><sup>HeLa cells transfected with a Cas9 expression vector (left) or untransfected cells (right) were fixed in methanol at -20°C, permeabilized with acetone at -20°C and blocked with PBS containing 2% BSA. The cells were stained with the Cas9 C-terminal antibody (Cat. No. C15200229) diluted 1:400, followed by incubation with an anti-mouse secondary antibody coupled to AF488. The bottom images show counter-staining of the nuclei with Hoechst 33342.</sup></p>
<h5><sup>Check our selection of antibodies validated in IF.</sup></h5>',
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'description' => '<p>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. They can undergo to multiple post-translational modifications (PTM), mainly at the N-terminal tails. These <strong>post-translational modifications </strong>include methylation, acetylation, phosphorylation, ubiquitinylation, citrullination, sumoylation, deamination and crotonylation. 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>”, 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 antibodies are highly specific and have been validated in many applications, including <strong>ChIP</strong> and <strong>ChIP-seq</strong>.</p>
<p>Diagenode’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’s highly validated antibodies:</span></p>
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<li><span style="font-weight: 400;"> Highly sensitive and specific</span></li>
<li><span style="font-weight: 400;"> Cost-effective (requires less antibody per reaction)</span></li>
<li><span style="font-weight: 400;"> Batch-specific data is available on the website</span></li>
<li><span style="font-weight: 400;"> Expert technical support</span></li>
<li><span style="font-weight: 400;"> Sample sizes available</span></li>
<li><span style="font-weight: 400;"> 100% satisfaction guarantee</span></li>
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<p><span style="font-weight: 400;">Diagenode provides leading solutions for epigenetic research. Because ChIP-seq is a widely-used technique, we validate our antibodies in ChIP and ChIP-seq experiments (in addition to conventional methods like Western blot, Dot blot, ELISA, and immunofluorescence) to provide the highest quality antibody. We standardize our validation and production to guarantee high product quality without technical bias. Diagenode guarantees ChIP-seq grade antibody performance under our suggested conditions.</span></p>
<div class="row">
<div class="small-12 medium-9 large-9 columns">
<p><strong>ChIP-seq profile</strong> of active (H3K4me3 and H3K36me3) and inactive (H3K27me3) marks using Diagenode antibodies.</p>
<img src="https://www.diagenode.com/img/categories/antibodies/chip-seq-grade-antibodies.png" /></div>
<div class="small-12 medium-3 large-3 columns">
<p><small> ChIP was performed on sheared chromatin from 100,000 K562 cells using iDeal ChIP-seq kit for Histones (cat. No. C01010051) with 1 µg of the Diagenode antibodies against H3K27me3 (cat. No. C15410195) and H3K4me3 (cat. No. C15410003), and 0.5 µg of the antibody against H3K36me3 (cat. No. C15410192). The IP'd DNA was subsequently analysed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along the complete sequence of human chromosome 3, a zoomin to a 10 Mb region and a further zoomin to a 1.5 Mb region. </small></p>
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<p>Diagenode’s highly validated antibodies:</p>
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<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> - 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> with rigorous QC and validation</li>
<li><strong>Classified</strong> 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. 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><span style="font-weight: 400;">All Diagenode’s antibodies are listed below. Please, use our Quick search field to find the antibody of interest by target name, application, purity.</span></p>
<p><span style="font-weight: 400;">Diagenode’s highly validated antibodies:</span></p>
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<li>Highly sensitive and specific</li>
<li>Cost-effective (requires less antibody per reaction)</li>
<li>Batch-specific data is available on the website</li>
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<p><br />Chromatin immunoprecipitation (<b>ChIP</b>) is a technique to study the associations of proteins with the specific genomic regions in intact cells. One of the most important steps of this protocol is the immunoprecipitation of targeted protein using the antibody specifically recognizing it. The quality of antibodies used in ChIP is essential for the success of the experiment. Diagenode offers extensively validated ChIP-grade antibodies, confirmed for their specificity, and high level of performance in ChIP. Each batch is validated, and batch-specific data are available on the website.</p>
<p></p>
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<p><strong>ChIP results</strong> obtained with the antibody directed against H3K4me3 (Cat. No. <a href="../p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>). </p>
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<div class="small-12 medium-6 large-6 columns"><img src="https://www.diagenode.com/img/product/antibodies/C15410003-fig1-ChIP.jpg" alt="" width="400" height="315" /> </div>
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<p>Our aim at Diagenode is to offer the largest collection of highly specific <strong>ChIP-grade antibodies</strong>. We add new antibodies monthly. Find your ChIP-grade antibody in the list below and check more information about tested applications, extensive validation data, and product information.</p>',
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'authors' => 'Ugai, Keita and Matsuda, Shuichi and Mikami, Hideki and Shimada, Ayako andMisawa, Tomoko and Nakamura, Hiroyuki and Tatsumi, Koichiro and Hatano,Masahiko and Murayama, Toshihiko and Kasuya, Yoshitoshi',
'description' => '<p>Idiopathic pulmonary fibrosis (IPF) is a fatal lung disease of unknown etiopathogenesis. The activation of extracellular matrix (ECM)-producing myofibroblasts plays a key role in fibrotic tissue remodeling. The dedifferentiation of myofibroblasts has attracted considerable attention as a promising target for the development of effective therapeutic interventions against IPF. Here, we screened a small library of epigenetics-related inhibitors using dedifferentiation assay of lung myofibroblasts prepared from a patient at the terminal stages of IPF and chose UNC0379. The inhibition of SET8, a histone H4 lysine 20 (H4K20) monomethyltransferase, by UNC0379 markedly suppressed the expression of α-smooth muscle actin (SMA) and ED-A-fibronectin in myofibroblasts. In IPF myofibroblasts, SET8 expression and H4K20 monomethylation (H4K20me1) levels, which were significantly higher than those in normal human lung fibroblasts, were reduced upon treatment with UNC0379. Hence, the changes in the expression of the two fibrotic markers clearly correlated with those in SET8 expression and H4K20me1 level. Furthermore, in a mouse model of bleomycin (BLM)-induced lung fibrosis, the intratracheal administration of UNC0379 at an early fibrotic stage markedly ameliorated the histopathological changes associated with collagen deposition in the lungs. However, treatment with UNC0379 did not significantly affect the number of proinflammatory cells or cytokine production in the bronchoalveolar lavage fluids from mice treated with BLM. In the BLM-injured lung, SET8 was predominantly localized to the nuclei of α-SMA-positive cells, which colocalized with H4K20me1. Taken together, our results indicate that the inhibition of {SET}8 resulting in myofibroblast dedifferentiation may partly mitigate lung fibrosis without affecting the inflammatory responses.</p>',
'date' => '2020-08-05',
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'authors' => 'Piazza R, Magistroni V, Redaelli S, Mauri M, Massimino L, Sessa A, Peronaci M, Lalowski M, Soliymani R, Mezzatesta C, Pirola A, Banfi F, Rubio A, Rea D, Stagno F, Usala E, Martino B, Campiotti L, Merli M, Passamonti F, Onida F, Morotti A, Pavesi F, Bregni',
'description' => '<p>SETBP1 variants occur as somatic mutations in several hematological malignancies such as atypical chronic myeloid leukemia and as de novo germline mutations in the Schinzel-Giedion syndrome. Here we show that SETBP1 binds to gDNA in AT-rich promoter regions, causing activation of gene expression through recruitment of a HCF1/KMT2A/PHF8 epigenetic complex. Deletion of two AT-hooks abrogates the binding of SETBP1 to gDNA and impairs target gene upregulation. Genes controlled by SETBP1 such as MECOM are significantly upregulated in leukemias containing SETBP1 mutations. Gene ontology analysis of deregulated SETBP1 target genes indicates that they are also key controllers of visceral organ development and brain morphogenesis. In line with these findings, in utero brain electroporation of mutated SETBP1 causes impairment of mouse neurogenesis with a profound delay in neuronal migration. In summary, this work unveils a SETBP1 function that directly affects gene transcription and clarifies the mechanism operating in myeloid malignancies and in the Schinzel-Giedion syndrome caused by SETBP1 mutations.</p>',
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'description' => 'DNA double-strand breaks (DSBs) activate a signaling pathway known as the DNA damage response (DDR) which via protein–protein interactions and post-translational modifications recruit signaling proteins, such as 53BP1, to chromatin flanking the lesion. Depletion of the SET8 methyltransferase prevents accumulation of 53BP1 at DSBs; however, this phenotype has been attributed to the role of SET8 in generating H4K20 methylation across the genome, which is required for 53BP1 binding to chromatin, prior to DNA damage. Here, we report that SET8 acts directly at DSBs during the DNA damage response (DDR). SET8 accumulates at DSBs and is enzymatically active at DSBs. Depletion of SET8 just prior to the induction of DNA damage abrogates 53BP1's accumulation at DSBs, suggesting that SET8 acts during DDR. SET8's occupancy at DSBs is regulated by histone deacetylases (HDACs). Finally, SET8 is functionally required for efficient repair of DSBs specifically via the non-homologous end-joining pathway (NHEJ). Our findings reveal that SET8's active role during DDR at DSBs is required for 53BP1's accumulation.',
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'name' => 'H4K20me1 Antibody',
'description' => '<p><span>Monoclonal antibody raised in mouse against histone <strong>H4 containing the monomethylated lysine 20 (H4K20me1)</strong>, using a KLH-conjugated synthetic peptide. </span></p>',
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--ChIP.png" alt="H4K20me1 Antibody ChIP Grade" /></p>
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<p><small><strong> Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />ChIP assays were performed using human HeLa cells, the Diagenode monclonal antibody against H4K20me1 (cat. No. C15200147) and optimized PCR primer sets for qPCR. ChIP was performed with the “the “iDeal ChIP-seq” kit (cat. No. C01010051), using sheared chromatin from 1 million cells. A titration of the antibody consisting of 0.5, 1, 2 and 5 μg per ChIP experiment was analysed. IgG (1 μg/IP) was used as negative IP control. QPCR was performed with primers for the coding region of the active GAPDH and ACTB genes, used as positive controls, and for the GAPDH promoter and a region located 1 kb upstream of the GAPDH promoter, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" /></p>
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<div class="small-7 columns">
<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-A.png" alt="H4K20me1 Antibody ChIP-seq Grade " caption="false" width="367" height="60" /></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-D.png" alt="H4K20me1 Antibody validated in ChIP-seq" caption="false" width="367" height="60" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
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<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" caption="false" width="280" height="180" /></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" caption="false" width="367" height="139" /></p>
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<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-B.png" alt="H4K20me1 Antibody for ChIP-seq" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-ChIPSeq-C.png" alt="H4K20me1 Antibody for ChIP-seq assay" /></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
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<p><small><strong> Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />ChIP was performed as described above using 0.5 μg of the Diagenode antibody against H4K20me1 (cat. No. C15200147). The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution in genomic regions from chromosomes 12, 7, 14 and 3 surrounding the GAPDH, ACTB, FOS and EIF4A2 positive control genes (figure 2A, B, C and D, respectively). </small></p>
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</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-DotBlot.png" alt="H4K20me1 Antibody Dot Blot validation" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 3. Cross reactivity tests using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />To check the specificity of the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure 3 shows a high specificity of the antibody for the modification of interest. </small></p>
</div>
</div>
<div class="row">
<div class="small-4 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147--WB.png" alt="H4K20me1 Antibody validated in Western Blot" /></p>
</div>
<div class="small-8 columns">
<p><small><strong> Figure 4. Western blot analysis using the Diagenode monoclonal antibody directed against H4K20me1 </strong><br />Western blot was performed on whole cell (25 μg, lane 1) and histone extracts (15 μg, lane 2) from HeLa cells, and on 1 μg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Diagenode monoclonal antibody against H4K20me1 (cat. No C15200147). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right </small></p>
</div>
</div>
<div class="row">
<div class="small-5 columns">
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-A.png" alt="H4K20me1 Antibody validated in Immunofluorescence" /></p>
<p><img src="https://www.diagenode.com/img/product/antibodies/C15200147-IF-B.png" alt="H4K20me1 Antibody validated for Immunofluorescence" /></p>
</div>
<div class="small-7 columns">
<p><small><strong> Figure 5. Immunofluorescence using the Diagenode monoclonal antibody directed against H4K20me1</strong> <br />Human osteosarcoma (U2OS) cells were stained with the Diagenode antibody against H4K20me1 (cat. No. MAb-147- 100) and with DAPI. Cells were fixed with ice cold methanol for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum. Figure 4A: cells were immunofluorescently labeled with the H4K20me1 antibody (left), diluted 1:200 in blocking solution followed by an anti- mouse antibody conjugated to Alexa488 or with DAPI (right), which specifically labels DNA. Figure 4B: staining of the cells with the H4K20me1 antibody after incubation of the antibody with blocking peptide (cat. No. sp-034-050, concentration: 5 ng/μl). </small></p>
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'name' => 'SET8 methyltransferase activity during the DNA double-strand break response is required for recruitment of 53BP1',
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'description' => 'DNA double-strand breaks (DSBs) activate a signaling pathway known as the DNA damage response (DDR) which via protein–protein interactions and post-translational modifications recruit signaling proteins, such as 53BP1, to chromatin flanking the lesion. Depletion of the SET8 methyltransferase prevents accumulation of 53BP1 at DSBs; however, this phenotype has been attributed to the role of SET8 in generating H4K20 methylation across the genome, which is required for 53BP1 binding to chromatin, prior to DNA damage. Here, we report that SET8 acts directly at DSBs during the DNA damage response (DDR). SET8 accumulates at DSBs and is enzymatically active at DSBs. Depletion of SET8 just prior to the induction of DNA damage abrogates 53BP1's accumulation at DSBs, suggesting that SET8 acts during DDR. SET8's occupancy at DSBs is regulated by histone deacetylases (HDACs). Finally, SET8 is functionally required for efficient repair of DSBs specifically via the non-homologous end-joining pathway (NHEJ). Our findings reveal that SET8's active role during DDR at DSBs is required for 53BP1's accumulation.',
'date' => '2014-09-25',
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