Cited by 306Receptor Mechanisms and SignalingMonoclonal and Polyclonal Antibodies ResearchNeuropeptides and Animal Physiology

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How reliable are G-protein-coupled receptor antibodies?

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Michel, Martin C., Wieland, Thomas, Tsujimoto, Gozoh·2009·Naunyn-Schmiedeberg's Archives of Pharmacology

DOI 10.1007/s00210-009-0395-y

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Abstract

A cluster of manuscripts in this issue of the Journal highlights a lack of selectivity of 49 antibodies against 19 subtypes of alpha(1)- and beta-adrenoceptors, muscarinic, dopamine and galanin receptors as well as vanilloid (TRPV1) receptors. Taken together these data demonstrate that lack of selectivity appears to be the rule rather than the exception for antibodies against G-protein-coupled and perhaps also other receptors. Thus, the previously often applied validation of such antibodies by the disappearance of staining in the presence of blocking peptide, i.e. the antigen against which the antibody was raised, alone is insufficient to demonstrate specificity. We propose that receptor antibodies should be validated by at least one of the following techniques: a) disappearance of staining in knock-out animals of the target receptor, b) reduction of staining upon knock-down approaches such as siRNA treatment, c) selectivity of staining in immunoblots or immunocytochemistry for the target receptor vs. related subtypes when expressed in the same cell line and/or d) antibodies raised against multiple distinct epitopes of a receptor yielding very similar staining patterns. Other issues of consideration to obtain reliable results based on receptor antibodies in applications such as immunohistochemistry or immunoblotting are also being discussed.

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Authors: Michel, M. C., Wieland, T., & Tsujimoto, G.
Affiliations: Amsterdam UMC Location University of Amsterdam (NL)
Heidelberg University (DE)
Kyoto University (JP)
Corresponding author: Michel, Martin C.
ORCID: Michel, Martin C.
Wieland, Thomas
Venue: Naunyn-Schmiedeberg's Archives of Pharmacology
Publisher: Springer Science and Business Media LLC
Published: 2009-01-27
Volume: 379
Issue: 4
Pages: 385-388
ISSN: 0028-1298 (ISSN-L)
1432-1912
DOI: 10.1007/s00210-009-0395-y
OpenAlex: W2029187954
Cited by: 306
References: 28
Citations by year: 2026: 2 · 2025: 11 · 2024: 18 · 2023: 19 · 2022: 19 · 2021: 23 · 2020: 16 · 2019: 19 · 2018: 14 · 2017: 21 · 2016: 14 · 2015: 30 · 2014: 23 · 2013: 18 · 2012: 21
FWCI: 7.74
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by-nc
Locations: Naunyn-Schmiedeberg s Archives of Pharmacology (publishedVersion, cc-by-nc)
PubMed (publishedVersion)
Pure Amsterdam UMC (publishedVersion)
UvA-DARE (University of Amsterdam) (acceptedVersion, other-oa)
Data Archiving and Networked Services (DANS) (submittedVersion)
Data Archiving and Networked Services (DANS) (submittedVersion, other-oa)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
Life Sciences › Immunology and Microbiology › Immunology › Monoclonal and Polyclonal Antibodies Research
Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuropeptides and Animal Physiology
Keywords: G protein-coupled receptors, antibody specificity, cross-reactivity, adrenergic receptors, muscarinic receptors, dopamine receptors, galanin receptors, TRPV1, blocking peptides, siRNA knockdown, epitope specificity, immunohistochemistry, Western blot
MeSH terms: Animals, Antibodies, Antibody Specificity, Humans, Receptors, Adrenergic, Receptors, Dopamine, Receptors, Muscarinic, Conserved Sequence, Sequence Homology, Amino Acid, Editorial Policies, Receptors, G-Protein-Coupled, Receptors, Galanin, TRPV Cation Channels, Validation Studies as Topic

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