westernblot鉴定精华中的精华BTN_A_000113382_O_88017a.pdf

Review Review Antibody validation Jennifer Bordeaux, Allison W. Welsh, Seema Agarwal, Elizabeth Killiam, Maria T. Baquero, Jason A. Hanna, Valsamo K. Anagnostou, and David L. Rimm Department of Pathology, Yale University School of Medicine, New Haven, CT, USA BioTechniques 48:197-209 (March 2010) doi 10.2144/000113382 Keywords: antibody; validation; immunohistochemistry; immunofluorescence 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. Taylor and David Dabbs, and recently, an For antibodies, one must demonstrate that Introduction ad hoc group published a set of “recom- they are specific, selective, and reproducible Antibodies are among the most commonly mendations” (2). However, these works in the context for which they are used. used research tools, routinely used for focus on the clinical aspects of IHC, When it comes to IHC, standardization Western blot (WB), immunoprecipit