Ten commercial antibodies for alpha-1-adrenergic receptor subtypes are nonspecific
- 557 Downloads
Commercial antibodies are used widely to quantify and localize the α1-adrenergic receptor (AR) subtypes, α1A, α1B, and α1D. We tested ten antibodies, from abcam and Santa Cruz, using western blot with heart and brain tissue from wild-type (WT) mice and mice with systemic knockout (KO) of one or all three subtypes. We found that none of the antibodies detected a band in WT that was absent in the appropriate KO or in the KO that was null for all α1-ARs (ABDKO). We conclude that the antibodies we tested are not specific for α1-ARs. These results raise caution with prior studies using these reagents. For now, competition radioligand binding is the only reliable approach to quantify the α1-AR subtype proteins. Receptor protein localization remains a challenge.
KeywordsAlpha-1-adrenoceptor Antibodies Subtypes
We appreciate the support of the United States Department of Veterans Affairs Research Service, the NIH, and the GlaxoSmithKline Research Foundation.
- Cavalli A, Lattion AL, Hummler E, Nenniger M, Pedrazzini T, Aubert JF, Michel MC, Yang M, Lembo G, Vecchione C, Mostardini M, Schmidt A, Beermann F, Cotecchia S (1997) Decreased blood pressure response in mice deficient of the alpha1b-adrenergic receptor. Proc Natl Acad Sci U S A 94:11589–11594PubMedCrossRefGoogle Scholar
- Jensen BC, Swigart PM, Laden M-E, DeMarco T, Hoopes C, Simpson PC (2008) The alpha-1D is the predominant alpha-1 adrenergic receptor subtype in human epicardial coronary arteries (abstract). Arterioscler Thromb Vasc Biol 28:e142Google Scholar
- O'Connell TD, Ishizaka S, Nakamura A, Swigart PM, Rodrigo MC, Simpson GL, Cotecchia S, Rokosh DG, Grossman W, Foster E, Simpson PC (2003) The alpha(1A/C)- and alpha(1B)-adrenergic receptors are required for physiological cardiac hypertrophy in the double-knockout mouse. J Clin Invest 111:1783–1791PubMedCrossRefGoogle Scholar
- Simpson PC (2006) Lessons from Knockouts: the alpha-1-ARs. In: Perez DM (ed) The adrenergic receptors in the 21st century. Humana, Totowa, pp 207–240Google Scholar