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Title: Lack of specificity of commercially available antisera: Better specifications needed
Authors: Pradidarcheep W.
Labruyère W.T.
Dabhoiwala N.F.
Lamers W.H.
Keywords: antiserum
muscarinic receptor
amino acid sequence
animal experiment
animal tissue
antibody affinity
antibody specificity
knockout mouse
priority journal
protein analysis
Western blotting
Amino Acid Sequence
Antibody Specificity
Blotting, Western
Gastrointestinal Tract
Immune Sera
Mice, Knockout
Molecular Sequence Data
Rats, Wistar
Receptors, Muscarinic
Sensitivity and Specificity
Urinary Tract
Issue Date: 2008
Abstract: The ideal antiserum for immunohistochemical (IHC) applications contains mono-3 specific high-affinity antibodies with little nonspecific adherence to sections. Many commercially available antibodies are "affinity" purified, but it is unknown if they meet "hard" specificity criteria, such as absence of staining in tissues genetically deficient for the antigen or a staining pattern that is identical to that of an antibody raised against a different epitope on the same protein. Reviewers, therefore, often require additional characterization. Although the affinity-purified antibodies used in our study on the distribution of muscarinic receptors produced selective staining patterns on sections, few passed the preabsorption test, and none produced bands of the anticipated size on Western blots. More importantly, none showed a difference in staining pattern on sections or Western blots between wild-type and knockout mice. Because these antibodies were used in most studies published thus far, our findings cast doubts on the validity of the extant body of morphological knowledge of the whole family of muscarinic receptors. We formulate requirements that antibody-specification data sheets should meet and propose that journals for which IHC is a core technique facilitate consumer rating of antibodies. "Certified" antibodies could avoid fruitless and costly validation assays and should become the standard of commercial suppliers. © The Histochemical Society, Inc.
ISSN: 221554
Appears in Collections:Scopus 1983-2021

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