ELISA was performed using a serial dilution of H3 polyclonal antibody (Cat # PAB14096). By plotting the absorbance against the antibody dilution, the titer of the crude serum was estimated to be 1 : 900,000.
A Dot Blot analysis was performed to test the cross reactivity of H3 polyclonal antibody (Cat # PAB14096) with the peptides used for immunization of the rabbit and other peptides containing different modifications or 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 : 1,000. Figure shows a high specificity of the crude serum for one of the unmodified peptides.