Publication:
Lack of specificity of commercially available antisera: Better specifications needed

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Issued Date

2008

Resource Type

Article

File Type

application/pdf

ISSN

221554

DOI

10.1369/jhc.2008.952101

Other identifier(s)

2-s2.0-56449096774

Rights Holder(s)

มหาวิทยาลัยศรีนครินทรวิโรฒ

Bibliographic Citation

Journal of Histochemistry and Cytochemistry. Vol 56, No.12 (2008), p.1099-1111

Suggested Citation

Pradidarcheep W., Labruyère W.T., Dabhoiwala N.F., Lamers W.H. Lack of specificity of commercially available antisera: Better specifications needed. Journal of Histochemistry and Cytochemistry. Vol 56, No.12 (2008), p.1099-1111. doi:10.1369/jhc.2008.952101 Retrieved from: https://hdl.handle.net/20.500.14740/3808

Title

Lack of specificity of commercially available antisera: Better specifications needed

Author(s)

Pradidarcheep W.
 Labruyère W.T.
 Dabhoiwala N.F.
 Lamers W.H.

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.

Subject(s)

Antiserum
 Epitope
 Muscarinic receptor
 Amino acid sequence
 Animal experiment
 Animal tissue
 Antibody affinity
 Antibody specificity
 Article
 Immunohistochemistry
 Knockout mouse
 Mouse
 Nonhuman
 Priority journal
 Protein analysis
 Rat
 Staining
 Western blotting
 Amino Acid Sequence
 Animals
 Antibody Specificity
 Blotting, Western
 Brain
 Epitopes
 Gastrointestinal Tract
 Immune Sera
 Immunohistochemistry
 Mice
 Mice, Knockout
 Molecular Sequence Data
 Rats
 Rats, Wistar
 Receptors, Muscarinic
 Sensitivity and Specificity
 Urinary Tract
 Mus

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URI

https://hdl.handle.net/20.500.14740/3808

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Scopus: Year 1983-2021

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