Antibody validation.
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.
Bordeaux
Jennifer
J
Department of Pathology, Yale University School of Medicine, New Haven, CT 06520, USA.
Welsh
Allison
A
Agarwal
Seema
S
Killiam
Elizabeth
E
Baquero
Maria
M
Hanna
Jason
J
Anagnostou
Valsamo
V
Rimm
David
D
D000465:Algorithms; D000818:Animals; D000906:Antibodies / Q000032:analysis*; D001709:Biotechnology / Q000592:standards*; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D000906:Antibodies