WB
Western blot analysis of various lysates using Acetyl-Histone H3-K14 Rabbit pAb at 1:10000 dilution incubated overnight at 4℃. HeLa cells were treated with TSA (1 uM) at 37°C for 18 hours, NIH/3T3 cells were treated with TSA (1uM) at 37°C for 18 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit.
Exposure time: 20 s.
CHIP
Chromatin immunoprecipitation was performed with 10 μg of cross-linked chromatin from HeLa cells, using 3 μg of Acetyl-Histone H3-K14 Rabbit pAb and Rabbit IgG isotype control. The enrichment of immunoprecipitated DNA at different genomic loci was examined by quantitative PCR. The histogram compares the ratio of the immunoprecipitated DNA to the input at given loci.