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

• Published in: Naunyn-Schmiedeberg's archives of pharmacology Vol. 379; no. 4; pp. 409 - 412
• Main Authors: Jensen, Brian C., Swigart, Philip M., Simpson, Paul C.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer-Verlag 01-04-2009
• Subjects: Animals; Antibodies - immunology; Antibody Specificity - immunology; Biomedical and Life Sciences; Biomedicine; Blotting, Western - methods; Brain Chemistry; Mice; Mice, Inbred C57BL; Mice, Knockout; Myocardium - chemistry; Neurosciences; Pharmacology/Toxicology; Receptors, Adrenergic, alpha-1 - analysis; Receptors, Adrenergic, alpha-1 - genetics; Receptors, Adrenergic, alpha-1 - immunology; Special Article; Antibodies; Alpha-1-adrenoceptor; Subtypes
• ISSN: 0028-1298; 1432-1912
• DOI: 10.1007/s00210-008-0368-6

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.