H4pan Polyclonal Antibody

HeLa cells were stained with H4pan Polyclonal Antibody (bs-53138R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 1% BSA. The cells were immunofluorescently labeled with the H4 antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

To determine the titer of the antibody, an ELISA was performed using a serial dilution of H4pan Polyclonal Antibody (bs-53138R) in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:3,000.

ChIP assays were performed using HeLa cells, H4pan Polyclonal Antibody (bs-53138R) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells on the IP-Star Compact automated system. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and c-fos genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).