Lack of specificity shown by P2Y6 receptor antibodies
Open accessAbstract
P2Y6 receptor in bladder smooth muscle responds to UDP by increasing muscle tone and augmenting bladder contractions. The exact cellular location of the receptor is however unknown. Three commercially available antibodies to P2Y6 receptor gave clean bands on Western blot which were eliminated by specific peptide competition. Two of the three also immunostained bladder smooth muscle cells while leaving adjacent interstitial cells of Cajal unstained. However, attempts to validate the specificity of these antibodies by performing the same assays on bladders from P2Y6 knockout mice were unsuccessful. In Western blots, all three antibodies bound similar proteins in both wild type and P2Y6 knockout tissue. Immunostaining of knockout tissue sections also showed no difference in staining patterns or intensity. We conclude that rigorous controls are required when using commercial reagents to this G-protein coupled receptor and perhaps to other members of the P2Y receptor family.
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By asking researchers to publish RRIDs, researchers were inadvertently encouraged to double-check their reagents, reducing errors in antibodies and the use of problematic cell lines, with no additional effort on the part of journals.
“By asking researchers to publish RRIDs, researchers were also inadvertently encouraged to double-check their reagents, reducing not only errors in antibodies but also the use of problematic cell lines, with no additional effort on the part of journals [29].”
From Open Science 2.0: Towards a truly collaborative research ecosystem (2023) · cites this paper as [29]
Simply citing a company name and antibody target leaves much ambiguity and, in a surprisingly large percentage of cases, leads scientists to waste money and time trying to optimize the wrong reagent.
“Simply citing a company name and target leaves much ambiguity and, in a surprisingly large percentage of cases, leads scientists to waste money and time trying to optimize the wrong reagent [27–29].”
From Open Science 2.0: Towards a truly collaborative research ecosystem (2023) · cites this paper as [29]
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- Authors
- Yu, W., & Hill, W. G.
- Affiliations
- Beth Israel Deaconess Medical Center (US)
- Corresponding author
- Hill, Warren G.
- Venue
- Naunyn-Schmiedeberg's Archives of Pharmacology
- Published
- 2013-06-22
- Volume
- 386
- Issue
- 10
- Pages
- 885-891
- ISSN
- 0028-1298 (ISSN-L)
- 1432-1912
- PMC
- PMC3772977
- OpenAlex
- W2030694434
- Cited by
- 36
- References
- 33
- Citations by year
- 2024: 1 · 2023: 2 · 2022: 2 · 2020: 1 · 2019: 2 · 2018: 5 · 2017: 3 · 2016: 2 · 2015: 8 · 2014: 10
- FWCI
- 5.54
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Naunyn-Schmiedeberg s Archives of Pharmacology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Institutes of Health
- National Institute of Diabetes and Digestive and Kidney Diseases (R00DK095922, R01DK083299)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Adenosine and Purinergic Signaling
- Health Sciences › Medicine › Urology › Urinary Bladder and Prostate Research
- Health Sciences › Medicine › Gastroenterology › Gastrointestinal motility and disorders
- Keywords
- P2Y6 receptor, antibody specificity, bladder smooth muscle, UDP, G protein-coupled receptors, interstitial cells of Cajal, receptor localization
- MeSH terms
- Animals, Antibodies, Urinary Bladder, Mice, Inbred C57BL, Muscle, Smooth, Receptors, Purinergic P2, Mice, Knockout, Mice, In Vitro Techniques
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