Ten commercial antibodies for alpha-1-adrenergic receptor subtypes are nonspecific.
Commercial antibodies are used widely to quantify and localize the alpha1-adrenergic receptor (AR) subtypes, alpha1A, alpha1B, and alpha1D. 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 alpha1-ARs (ABDKO). We conclude that the antibodies we tested are not specific for alpha1-ARs. These results raise caution with prior studies using these reagents. For now, competition radioligand binding is the only reliable approach to quantify the alpha1-AR subtype proteins. Receptor protein localization remains a challenge.
Jensen
Brian C
BC
Cardiology Section and Research Service, San Francisco VA Medical Center (111-C-8), 4150 Clement St., San Francisco, CA 94121, USA.
Swigart
Philip M
PM
Simpson
Paul C
PC
D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western / Q000379:methods; D001923:Brain Chemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009206:Myocardium / Q000737:chemistry; D018340:Receptors, Adrenergic, alpha-1 / Q000032:analysis* / Q000235:genetics / Q000276:immunology*
C547714:Adra1a protein, mouse; C547717:Adra1b protein, mouse; C547720:Adra1d protein, mouse; D000906:Antibodies; D018340:Receptors, Adrenergic, alpha-1