ELISA was performed using a serial dilution of H4K8ac polyclonal antibody (Cat # PAB14070). 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 : 17,500.
A Dot Blot analysis was performed to test the cross reactivity of H4K8ac polyclonal antibody (Cat # PAB14070) with other modifications of histone H4 and H3 and with the unmodified histone H4. 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 : 20,000. Figure shows a high specificity of the crude serum for the modification of interest.