Western blot analysis of extracts of various cell lines, using Anti-CTCF Antibody (A12562) at 1:500 dilution. Secondary antibody: Goat Anti-Rabbit IgG (H+L) (HRP) (AS014) at 1:10,000 dilution. Lysates / proteins: 25µg per lane. Blocking buffer: 3% non-fat dry milk in TBST. Detection: ECL Basic Kit (RM00020). Exposure time: 5s.
Chromatin immunoprecipitation of extracts of HCT116 cell line, using Anti-CTCF Antibody (A12562) 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.