Enhanced validation of antibodies for research applications. Issue 1 (December 2018)

• Record Type: Journal Article
• Authors: Edfors, Fredrik; Hober, Andreas; Linderbäck, Klas; Maddalo, Gianluca; Azimi, Alireza; Sivertsson, Åsa; Tegel, Hanna; Hober, Sophia; Szigyarto, Cristina; Fagerberg, Linn; Feilitzen, Kalle; Oksvold, Per; Lindskog, Cecilia; Forsström, Björn; Uhlen, Mathias
• Is Part Of: Nature communications. Volume 9:Issue 1(2018)
• Page Start: 1
• Page End: 10
• Publication Date: 2018-12
• Subjects: Biology -- Periodicals; Physical sciences -- Periodicals; 505
• DOI: 10.1038/s41467-018-06642-y
• Languages: English
• ISSNs: 2041-1723

Abstract

Abstract There is a need for standardized validation methods for antibody specificity and selectivity. Recently, five alternative validation pillars were proposed to explore the specificity of research antibodies using methods with no need for prior knowledge about the protein target. Here, we show that these principles can be used in a streamlined manner for enhanced validation of research antibodies in Western blot applications. More than 6, 000 antibodies were validated with at least one of these strategies involving orthogonal methods, genetic knockdown, recombinant expression, independent antibodies, and capture mass spectrometry analysis. The results show a path forward for efforts to validate antibodies in an application-specific manner suitable for both providers and users. Five validation pillars have been proposed to verify the specificity of research antibodies. Here the authors screen 6, 000 antibodies from the Human Protein Atlas with these methods to provide an antibody validation resource for providers and users.