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Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K9me3 ChIP was performed with the Diagenode antibody against H3K9me3 (cat. No. C15210014) on sheared chromatin from 500,000 HeLaS3 cells using the “iDeal ChIP-seq” kit (cat. No. C01010051). A titration of the antibody consisting of 0.5, 1, and 2 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. Quantitative PCR was performed with primers for the ZNF510 gene and the Sat2 satellite repeat, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The graph shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against H3K9me3 ChIP was performed on sheared chromatin from 500,000 HeLaS3 cells using 1 µg of the Diagenode antibody against H3K9me3 (cat. No. C15210014) as described above. The IP'd DNA was subsequently analysed on an Illumina NovaSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2A shows the signal distribution along the long arm of chromosome 19 and a zoomin to an enriched region containing several ZNF repeat genes. The arrows indicate two satellite repeat regions which exhibit a stronger signal. Figures 2B, C and D show the enrichment genomic regions surrounding the ZNF510 positive control target and at the KCNQ1and H19 imprinted genes, respectively.
Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3K9me3 Western blot was performed on whole cell extracts (40 µg) from HeLa cells using the Diagenode antibody against H3K9me3 (cat. No. C15210014). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown on the right, the marker (in kDa) is shown on the left.
WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.
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