IMR Press / FBE / Volume 1 / Issue 1 / DOI: 10.2741/E21

Frontiers in Bioscience-Elite (FBE) is published by IMR Press from Volume 13 Issue 2 (2021). Previous articles were published by another publisher on a subscription basis, and they are hosted by IMR Press on imrpress.com as a courtesy and upon agreement with Frontiers in Bioscience.

Open Access Article

Detecting the mu opioid receptor in brain following SDS-PAGE with multiple approaches

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1 Department of Pharmacology and Center for Substance Abuse Research, Temple University School of Medicine, Philadelphia, PA 19140, USA
Academic Editor:Yingxian Pan
Front. Biosci. (Elite Ed) 2009, 1(1), 220–227; https://doi.org/10.2741/E21
Published: 1 June 2009
(This article belongs to the Special Issue regulation and function of opioid receptor genes)
Abstract

In general, it has been difficult to obtain antibodies which are useful for immunoblotting of endogenous seven-transmembrane receptors (7TMRs) despite the claims made by many companies on commercially available antibodies. In this review, we will use the mu opioid receptor (MOPR) in brain as an example to underscore the importance of using knock-out (K/O) mice and multiple independent approaches (ligand affinity-labeling, receptor phosphorylation and immunoblotting) in identifying 7TMRs following sodium dodecyl sulfate – polyacrylamide gel electrophoresis (SDS-PAGE). The rigor and convergence of pharmacological and biochemical data provide confidence on the unequivocal identification of MOPR. The distinct relative molecular masses (Mr's) and band patterns are largely due to variations in the extent of N-glycosylation in different cell lines, brain regions and species.

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