WB
Western blot analysis of various lysates using Acetyl-Histone H4-K5 Rabbit pAb at 1:1000 dilution. HeLa cells,NIH/3T3 cells and C6 cells were treated with TSA (1 uM) at 37℃ for 18 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit.
Exposure time: 60s.
CHIP
Chromatin immunoprecipitation analysis of extracts of HeLa cells, using Acetyl-Histone H4-K5 antibody and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.