Edfors, F. et al. Nat. Commun. 9, 4130 (2018).
The need for better validation of the specificity and reproducibility of antibody reagents for biological research applications has been a recent hot-button topic. To begin to address the many challenges, the International Working Group for Antibody Validation recently proposed five ‘pillars’ for antibody validation: genetic knockdown, recombinant expression, independent antibodies, capture mass spectrometry analysis, and orthogonal methods. Edfors et al. applied the five pillars, including an original approach for orthogonal validation combining transcriptomics and proteomics analysis across multiple cell lines, to thousands of antibodies in the Human Protein Atlas resource in order to validate their use in western blotting. They report more than 6,000 antibodies with enhanced validation, meaning that their good performance for western blotting was validated by at least one of the pillar approaches; 263 antibodies were validated by three or more pillars.
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Doerr, A. Enhanced antibody validation. Nat Methods 15, 1001 (2018). https://doi.org/10.1038/s41592-018-0248-z