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

Naunyn Schmiedebergs Arch Pharmacol. 2009 Apr;379(4):409-12. doi: 10.1007/s00210-008-0368-6. Epub 2008 Nov 7.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Validation Study

MeSH terms

  • Animals
  • Antibodies / immunology*
  • Antibody Specificity / immunology*
  • Blotting, Western / methods
  • Brain Chemistry
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Myocardium / chemistry
  • Receptors, Adrenergic, alpha-1 / analysis*
  • Receptors, Adrenergic, alpha-1 / genetics
  • Receptors, Adrenergic, alpha-1 / immunology*

Substances

  • Adra1a protein, mouse
  • Adra1b protein, mouse
  • Adra1d protein, mouse
  • Antibodies
  • Receptors, Adrenergic, alpha-1