Do you perform western blot testing of your histone antibodies on whole cell extracts or purified histones?

We use whole cell extracts when validating and QC testing our histone antibodies by western blot and do not routinely test purified histones. We generally load 20-30ug of whole cell extracts on the gel but have seen good signal with less than 20ug for some of our stronger histone antibodies. Since we do not routinely test purified histones we do not have a specific amount we can recommend. We suggest titrating your purified histones to determine the optimal amount to use for your specific antibody. Most of our histone antibodies are very strong, but this should be empirically determined for your specific samples.

Last updated: September 12, 2024