ELISA was performed using a serial dilution of H3K79me2 polyclonal antibody (Cat # PAB14047). 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 : 4,500.
A Dot Blot analysis was performed to test the cross reactivity of H3K79me2 polyclonal antibody (Cat # PAB14047) with other modifications of histone H3. Other H3 modifications include mono- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 36. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1 : 50,000. Figure shows a high specificity of the crude serum for the modification of interest.