Ten commercial antibodies for alpha-1-adrenergic receptor subtypes are nonspecific

Commercial antibodies are used widely to quantify and localize the alpha 1-adrenergic receptor (AR) subtypes, alpha 1A, alpha 1B, and alpha 1D. 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 alpha 1-ARs (ABDKO). We conclude that the antibodies we tested are not specific for alpha 1-ARs. These results raise caution with prior studies using these reagents. For now, competition radioligand binding is the only reliable approach to quantify the alpha 1-AR subtype proteins. Receptor protein localization remains a challenge.