ELISA was performed using a serial dilution of H3K27me1 polyclonal antibody (Cat # PAB14115), 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, the titer of the purified antibody was estimated to be 1 : 11,900.
A Dot Blot analysis was performed to test the cross reactivity of H3K27me1 polyclonal antibody (Cat # PAB14115) with peptides containing other modifications of histone H3 and the unmodified H3K27. 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 3 shows a high specificity of the antibody for the modification of interest.