WB
Western blot analysis of lysates from HeLa cells, using Acetyl-Histone H3-K56 Rabbit pAb at 1:10000 dilution. HeLa 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: 30s.
WB
Western blot analysis of lysates from NIH/3T3 cells, using Acetyl-Histone H3-K56 Rabbit pAb at 1:10000 dilution. NIH/3T3 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: 30s.
ICC/IF
Immunofluorescence analysis of C6 treated with TSA jia C6 cells using Acetyl-Histone H3-K56 Rabbit pAb at dilution of 100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
ICC/IF
Immunofluorescence analysis of U-2 OS treated with TSA jia U-2 OS cells using Acetyl-Histone H3-K56 Rabbit pAb at dilution of 100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) at 1:500 dilution. Blue: DAPI for nuclear staining.
CHIP
Chromatin immunoprecipitation analysis of extracts of Hale cells, using Acetyl-Histone H3-K56 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.