Histone H2B [ac Lys12] Antibody

Western Blot: Histone H2B [ac Lys12] Antibody [NBP2-59271] - Histone H2B Antibody [NBP2-59271] - Western blot was performed on whole cell (25 ug, lane 1) and histone extracts (15 ug, lane 2) from HeLa cells, and on 1 ug of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the antibody against H2BK12ac. The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (kDa) is shown on the left.

Chromatin Immunoprecipitation: Histone H2B [ac Lys12] Antibody [NBP2-59271] - Histone H2B Antibody [NBP2-59271] - ChIP assays were performed using human HeLa cells, the antibody against H2BK12ac and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH and ACTB promoters, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ELISA: Histone H2B [ac Lys12] Antibody [NBP2-59271] - Histone H2B Antibody [NBP2-59271] - To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against H2BK12ac in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:38,200.