Assay of GTPγS Binding in Autoradiography

Methods Mol Biol. 2016:1412:95-101. doi: 10.1007/978-1-4939-3539-0_10.

Abstract

Autoradiography of radiolabeled GTPγS ([(35)S]GTPγS) binding is a relevant method to study the function of G protein-coupled receptors (GPCRs), in tissue sections. Here, we describe the protocol for such a binding autoradiography, suitable to investigate the functionality of CB1 receptor in tissue slices from rodent brain.

Keywords: Autoradiography; Binding; Brain section; CB1 receptor; G-proteins; [35S]GTPγS.

MeSH terms

  • Animals
  • Autoradiography* / methods
  • Brain / metabolism
  • Guanosine 5'-O-(3-Thiotriphosphate) / metabolism*
  • Isotope Labeling*
  • Mice
  • Rats
  • Receptor, Cannabinoid, CB1 / agonists
  • Receptor, Cannabinoid, CB1 / metabolism
  • Sulfur Radioisotopes*

Substances

  • Receptor, Cannabinoid, CB1
  • Sulfur Radioisotopes
  • Guanosine 5'-O-(3-Thiotriphosphate)