H2Apan Polyclonal Antibody

HeLa cells were stained with H2Apan Polyclonal Antibody (bs-53140R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H2Apan antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle 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 H2Apan Polyclonal Antibody (bs-53140R) in antigen-coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:32,500.

ChIP assays were performed using HeLa cells, \r\nH2Apan Polyclonal Antibody (bs-53140R) and optimized PCR primer sets for qPCR. ChIP was performed with sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).