Abstract
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.
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We appreciate the support of the United States Department of Veterans Affairs Research Service, the NIH, and the GlaxoSmithKline Research Foundation.
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Jensen, B.C., Swigart, P.M. & Simpson, P.C. Ten commercial antibodies for alpha-1-adrenergic receptor subtypes are nonspecific. Naunyn-Schmied Arch Pharmacol 379, 409–412 (2009). https://doi.org/10.1007/s00210-008-0368-6
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DOI: https://doi.org/10.1007/s00210-008-0368-6