ELISA was performed using a serial dilution of H3K9me3S10p polyclonal antibody (Cat # PAB14091). 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 : 87,000.
A Dot Blot analysis was performed to test the cross reactivity of H3K9me3S10p polyclonal antibody (Cat # PAB14091) with peptides containing other modifications and unmodified sequences of histone H3. 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 : 1,000. Figure shows a high specificity of the crude serum for the modification of interest.