ELISA was performed using a serial dilution of H3K36me1 polyclonal antibody (Cat # PAB14132), crude serum and Flow Through. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the purified antibody was estimated to be 1 : 14,000.
A Dot Blot analysis was performed to test the cross reactivity of H3K36me1 polyclonal antibody (Cat # PAB14132) with peptides containing other modifications and unmodified sequences of histone H3. Other H3 modifications include di- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1 : 10,000. Figure shows a slight cross reaction of the antibody with the dimethylated H3K36.