Knockout validation as standard antibody validation
One of the most accepted and trusted validation processes for antibody specificity is through knockout validation (Madhusoodanan, 2014; Rhodes & Trimmers, 2008). In a knockout validation, antibody specificity is confirmed by testing the antibody of interest on a knockout sample or cell line which doesn't express the target protein. In a knockout cell line, the antibody target is not present as the gene encoding the protein of interest is eliminated or "knocked out". This data is compared side-by-side against a normal cell line (or wild type) and if the antibody is truly specific, the antibody should have no detection in the knockout cell line, but instead a specific signal in the normal cell line. Knockout validations offers a true negative control to confirm the antibody specificity to the protein of interest (Bordeaux, et al., 2010).
See all the knockout validation data below