Antibodies are among the most frequently used tools in research and have had a profound impact on the discovery of diagnostic and therapeutic targets and the understanding of the molecular background of diseases. In particular in reverse phase protein arrays (RPPA), where there is no separation of the proteins according to molecular weight, it is crucial that antibodies are proven to be highly specific, selective, and reproducible. However, numerous studies have shown that many antibodies frequently do not recognize the protein that they are supposed to detect, that multiple antibodies do often function in one application but not in another, and that antibodies are not stable over time or between different batches. So far, no universally accepted guidelines or standardized methods for determining the validity of antibodies have been established. This chapter discusses the urgent need for antibody validation, current strategies that are used for (RPPA) antibody validation, as well as proposes a new strategy about how to report, score, and integrate antibody validation from multiple users.
Keywords: Antibody; Proteomics; Reverse phase protein array (RPPA); Screening; Validation.