Antibody validation. (March 2010)

• Record Type: Journal Article
• Authors: Bordeaux, Jennifer; Welsh, Allison W.; Agarwal, Seema; Killiam, Elizabeth; Baquero, Maria T.; Hanna, Jason A.; Anagnostou, Valsamo K.; Rimm, David L.
• Is Part Of: Biotechniques. Volume 48:Number 3(2010)
• Page Start: 197
• Page End: 209
• Publication Date: 2010-03
• Subjects: antibody -- validation -- immunohistochemistry -- immunofluorescence; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000113382
• Languages: English
• ISSNs: 0736-6205

Abstract

Antibodies are among the most frequently used tools in basic science research and in clinical assays, but there are no universally accepted guidelines or standardized methods for determining the validity of these reagents. Furthermore, for commercially available antibodies, it is clear that what is on the label does not necessarily correspond to what is in the tube. To validate an antibody, it must be shown to be specific, selective, and reproducible in the context for which it is to be used. In this review, we highlight the common pitfalls when working with antibodies, common practices for validating antibodies, and levels of commercial antibody validation for seven vendors. Finally, we share our algorithm for antibody validation for immunohistochemistry and quantitative immunofluorescence.