ChIP assays were performed using the U-2 OS (human osteosarcoma cell line), Dnmt3b polyclonal antibody (Cat # PAB14125) and optimized PCR primer sets. Chromatin sheared from 1x106 cells and 0.2 ug of Dnmt3b antibody or beads only were used per ChIP experiment. The IP'd DNA was analysed at two loci : the MLH1 promoter, which is a positive locus for Dnmt3b, and the CDC6 promoter, a negative locus for Dnmt3b. Figure shows the recovery expressed as a % of the input DNA. Upper : Recovery of the MLH1 promoter by the Dnmt3b antibody or beads only. Bottom : Recovery of the CDC6 promoter by the Dnmt3b antibody or beads only.
ELISA was performed using a serial dilution of Dnmt3b polyclonal antibody (Cat # PAB14125) and crude serum in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1 : 900.