Cooper, T.McMurchie, E.Leifert, W.Lundstrom, K.Chiu, M.2010-03-302010-03-302009Methods in Molecular Biology, 2009; 552:143-15197816032731691064-37451940-6029http://hdl.handle.net/2440/57213The [³⁵S]GTPγS binding assay to measure G protein activation following agonist binding to G protein-coupled receptors (GPCRs) remains a powerful molecular technique to substantiate traditional pharmacological values of potency, efficacy, and affinity. The method described uses membrane preparations of the α2A-adrenergic receptor and purified G protein subunits expressed in Sf9 cells, reconstituted into a functional signaling system. This technology is generic and could be used with other GPCRs to demonstrate initial signaling events following receptor activation. Agonist-stimulated [³⁵S]GTPγS binding is measured in a 96-well plate format using scintillation counting.enCopyright 2009 Springer[35S]GTPγSα2A-adrenergic receptorG protein activationJournal article002009145810.1007/978-1-60327-317-6_102-s2.0-6765039203338319