Nuclear extracts of HeLa cells (15 ug) were analysed by Western blot using H3K4me2 polyclonal antibody (Cat # PAB14111) diluted 1 : 1,000 in TBS-Tween containing 5% skimmed milk.
ELISA was performed using a serial dilution of H3K4me2 polyclonal antibody (Cat # PAB14111), crude serum and flow through 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 : 18,300.
A Dot Blot analysis was performed to test the cross reactivity of H3K4me2 polyclonal antibody (Cat # PAB14111) with peptides containing other modifications and unmodified sequences of histone H3. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1 : 20,000. Figure shows a high specificity of the antibody for the modification of interest.