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

  • Brian C. Jensen
  • Philip M. Swigart
  • Paul C. SimpsonEmail author
Special Article


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.


Alpha-1-adrenoceptor Antibodies Subtypes 



We appreciate the support of the United States Department of Veterans Affairs Research Service, the NIH, and the GlaxoSmithKline Research Foundation.


  1. Cavalli A, Lattion AL, Hummler E, Nenniger M, Pedrazzini T, Aubert JF, Michel MC, Yang M, Lembo G, Vecchione C, Mostardini M, Schmidt A, Beermann F, Cotecchia S (1997) Decreased blood pressure response in mice deficient of the alpha1b-adrenergic receptor. Proc Natl Acad Sci U S A 94:11589–11594PubMedCrossRefGoogle Scholar
  2. Hein P, Michel MC (2007) Signal transduction and regulation: are all alpha1-adrenergic receptor subtypes created equal. Biochem Pharmacol 73:1097–1106PubMedCrossRefGoogle Scholar
  3. Jensen BC, Swigart PM, Myagmar B-E, Shah S, DeMarco T, Hoopes C, Simpson PC (2007) The alpha-1A is the predominant alpha-1-adrenergic receptor in the human heart at the mRNA but not the protein level (abstract). Circulation 116:II–289CrossRefGoogle Scholar
  4. Jensen BC, Swigart PM, Laden M-E, DeMarco T, Hoopes C, Simpson PC (2008) The alpha-1D is the predominant alpha-1 adrenergic receptor subtype in human epicardial coronary arteries (abstract). Arterioscler Thromb Vasc Biol 28:e142Google Scholar
  5. Koshimizu TA, Tanoue A, Tsujimoto G (2007) Clinical implications from studies of alpha1 adrenergic receptor knockout mice. Biochem Pharmacol 73:1107–1112PubMedCrossRefGoogle Scholar
  6. O'Connell TD, Ishizaka S, Nakamura A, Swigart PM, Rodrigo MC, Simpson GL, Cotecchia S, Rokosh DG, Grossman W, Foster E, Simpson PC (2003) The alpha(1A/C)- and alpha(1B)-adrenergic receptors are required for physiological cardiac hypertrophy in the double-knockout mouse. J Clin Invest 111:1783–1791PubMedCrossRefGoogle Scholar
  7. O'Connell TD, Swigart PM, Rodrigo MC, Ishizaka S, Joho S, Turnbull L, Tecott LH, Baker AJ, Foster E, Grossman W, Simpson PC (2006) Alpha1-adrenergic receptors prevent a maladaptive cardiac response to pressure overload. J Clin Invest 116:1005–1015PubMedCrossRefGoogle Scholar
  8. Rokosh DG, Simpson PC (2002) Knockout of the alpha 1A/C-adrenergic receptor subtype: the alpha 1A/C is expressed in resistance arteries and is required to maintain arterial blood pressure. Proc Natl Acad Sci U S A 99:9474–9479PubMedCrossRefGoogle Scholar
  9. Sadalge A, Coughlin L, Fu H, Wang B, Valladares O, Valentino R, Blendy JA (2003) alpha 1d Adrenoceptor signaling is required for stimulus induced locomotor activity. Mol Psychiatry 8:664–672PubMedCrossRefGoogle Scholar
  10. Simpson PC (2006) Lessons from Knockouts: the alpha-1-ARs. In: Perez DM (ed) The adrenergic receptors in the 21st century. Humana, Totowa, pp 207–240Google Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Brian C. Jensen
    • 1
    • 2
  • Philip M. Swigart
    • 1
  • Paul C. Simpson
    • 1
    • 3
    Email author
  1. 1.Cardiology Section and Research ServiceSan Francisco VA Medical Center (111-C-8)San FranciscoUSA
  2. 2.Cardiology Division, Department of MedicineUniversity of California, San Francisco (UCSF)San FranciscoUSA
  3. 3.Cardiovascular Research Institute, Department of MedicineUCSFSan FranciscoUSA

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