ELISA was performed using a serial dilution of H3K9ac polyclonal antibody (Cat # PAB14098) 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 purified antibody was estimated to be 1 : 6,100.
A Dot Blot analysis was performed to test the cross reactivity of H3K9ac polyclonal antibody (Cat # PAB14098) with peptides containing other histone modifications and the unmodified H3K9 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1 : 2,000. Figure shows a high specificity of the antibody for the modification of interest.