H2BK20ac Antibody - ChIP-seq Grade

Figure 1. ChIP results obtained with the Diagenode monoclonal antibody directed against H2BK20ac
ChIP assays were performed using human HeLa cells, the Diagenode antibody against H2BK20ac (cat. No. C15210010) 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 the active EIF4A2 and ACTB 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 H2BK20ac
ChIP was performed with 0.5 µg of the Diagenode antibody against H2BK20ac (cat. No. C15210010) 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 1.5 Mb region of the human X chromosome (figure 2A and B) and in two genomic regions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).

Figure 3. Western blot analysis using the Diagenode monoclonal antibody directed against H2BK20ac
Whole cell extracts from HeLa cells were analysed by Western blot using the Diagenode monoclonal antibody against H2BK20ac (cat. No. C15210010) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Figure 4. Immunofluorescence using the Diagenode monoclonal antibody directed against H2BK20ac
HeLa cells treated with sodium butyrate were stained with the Diagenode antibody against H2BK20ac No. C15210010, red) diluted 1:200. Actin filaments were stained with fluorescein phalladoin (green).