This chart shows an outline for initial antibody validation as published in Bordeaux et al; Biotechniques, 2010. Further testing of antibody specificity can be performed by transfecting cell lines that are known to be negative controls with the protein of interest and/or knockout cell lines expressing the biomarker of interest using siRNA or shRNA knockdown. Cell lines controls are formalin fixed and paraffin embedded and then further tested for antibody performance in FFPE blocks.
Bordeaux et al; Biotechniques. 2010 Mar;48(3):197-209.