ChIP assays were performed using human osteosarcoma (U-2 OS) cells, the polyclonal antibody against H3R17me2(asym) (Cat # PAB14059) and optimized PCR primer sets for qPCR. Figure A: qPCR performed with primers for the GAPDH promoter and for exon 2 of the myoglobin gene. Figure B: qPCR performed with primers for the promoter of the active ALDOA gene and for the coding region of the inactive MYOD gene.
ELISA was performed using a serial dilution of H3R17me2 polyclonal antibody (Cat # PAB14059). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the crude serum was estimated to be 1 : 40,000.