Mant-GTPγS triammonium

GTPγS-induced Inhibition of Binding

GTPγS-induced Inhibition of Binding 1. Chemicals and Equipment DB Buffer, PB, cAMP, cAMP [2, 8- 3H] (25 Ci/mmol, 1.0 mCi/ml, 32, 000 nM), 10 mM GTPγS, Wacker silicone AR-20, Reagiergefab test tubes (Sarstedt), millipore

Measurement of Agonist-Stimulated [35S]GTPγS Binding to Assess Total G-Protein and Gα-Subtype-Specific Activation by G-Protein-Coupled Receptors

preparations, GTP can be substituted by 35 S-labeled guanosine 5′-O -(3-thio)triphosphate ([35 S]GTγS) and on agonist stimulation a stable [35 S]GTPγS-Gα complex will form and accumulate. Separation of 35 S-bound GTPγS-Gα complexes from free [35 S]GTPγS allows

Cannabinoid Receptor Binding to Membrane Homogenates and Cannabinoid-Stimulated [35S]GTP-S Binding to Membrane Homogenates or Intact Cultured Cells

with it. Binding assays that assess the effect of an unlabeled receptor ligand on the binding of the radiolabeled guanosine triphosphate (GTP) analog [35 S]GTPγS provide such information and may be the most sensitive assays available for determining the relative