Semmelweis University Department of Medical Biochemistry
Unlike Western blots, RPPA does not resolve protein fractions by molecular weight, thus, signal could originate from unspecific primary and/or secondary antibody binding. Therefore, the reliability of this technique critically depends on antibody monospecificity. Antibodies are first validated for monospecificity and performance by conventional Western blotting, followed by RPPA. A Pearson correlation of >0.7 between WB and RPPA data from the same samples deems an antibody 'valid for RPPA'.