ELISA was performed using a serial dilution of H3K4me1 polyclonal antibody (Cat # PAB14112), 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 : 34,500.
A Dot Blot analysis was performed to test the cross reactivity of H3K4me1 polyclonal antibody (Cat # PAB14112) 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.