title: Western Blotting of the Endocannabinoid System.
abstract: Measuring expression levels of G protein-coupled receptors (GPCRs) is an important step for understanding the distribution, function, and regulation of these receptors. A common approach for detecting proteins from complex biological systems is Western blotting. In this chapter, we describe a general approach to Western blotting protein components of the endocannabinoid system using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nitrocellulose membranes, with a focus on detecting type 1 cannabinoid (CB1) receptors. When this technique is carefully used, specifically with validation of the primary antibodies, it can provide quantitative information on protein expression levels. Additional information can also be inferred from Western blotting such as potential posttranslational modifications that can be further evaluated by specific analytical techniques.
authors.affiliation: Department of Psychological and Brain Sciences, Indiana University, Bloomington, IN, 47405, USA.
authors.forename: Jim
authors.initials: J
authors.lastname: Wager-Miller
authors.affiliation: Department of Psychological and Brain Sciences, Indiana University, Bloomington, IN, 47405, USA. kmackie@indiana.edu.
authors.forename: Ken
authors.initials: K
authors.lastname: Mackie
mesh_terms: D000818:Animals; D015153:Blotting, Western / Q000379:methods; D063388:Endocannabinoids / Q000378:metabolism*; D006801:Humans; D043884:Receptor, Cannabinoid, CB1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D063388:Endocannabinoids; D043884:Receptor, Cannabinoid, CB1
keywords: Antibody; Electrophoresis; Electrophoretic transfer; Fluorescent detection