To determine the cross reactivity of H3K79me3 polyclonal antibody (Cat # PAB14050) with other histone H3 modifications, a Dot Blot analysis was performed. Tested histone H3 modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 36. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane and detected with the antibody diluted 1 : 50,000. Figure shows a high specificity for the modification of interest.
ELISA was performed using a serial dilution of H3K79me3 polyclonal antibody (Cat # PAB14050) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1 : 70,000.