Antibody validation

作者：Jennifer Bordeaux, Allison W. Welsh, Seema Agarwal, Elizabeth Killiam, Maria T. Baquero, Jason A. Hanna, Valsamo Anagnostou, David L. Rimm · 发表于：BioTechniques · 年份：2010 · DOI：10.2144/000113382 · 被引用次数：606 · 研究领域：Monoclonal and Polyclonal Antibodies Research、Advanced Biosensing Techniques and Applications、HER2/EGFR in Cancer Research

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.