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Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H3K23me2 ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3K23me2 (cat. No. C15210007) and optimized PCR primer sets for qPCR. ChIP was performed with the "iDeal ChIP-seq" kit (cat. No. C01010055) on sheared chromatin from 1,000,000 cells. A titration of the antibody consisting of 0.5, 1 and 2.5 µg per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for a region upstream of the promoters of the ACTB and GAPDH genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat region 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).
Figure 2. ChIP-seq results obtained with the Diagenode monoclonal antibody directed against H3K23me2 ChIP was performed with 0.5 µg of the Diagenode antibody against H3K23me2 (cat. No. C15210007) on sheared chromatin from 1,000,000 HeLa cells using the iDeal ChIP-seq kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the peak distribution along the complete sequence and a 3 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the ACTB and GAPDH positive control genes (figure 2C and D).
Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H3K23me2 Whole cell extracts from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H3K23me2 (cat. No. C15210007) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
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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