Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/57213
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Type: Book chapter
Title: [³⁵S]GTPγS binding in G protein-coupled receptor assays
Other Titles: [(35)S]GTPgammaS binding in G protein-coupled receptor assays
Author: Cooper, T.
McMurchie, E.
Leifert, W.
Citation: G Protein-coupled receptors in drug discovery, 2009 / Lundstrom, K., Chiu, M. (ed./s), vol.552, pp.143-151
Publisher: Humana Press
Publisher Place: United States
Issue Date: 2009
Series/Report no.: Methods in Molecular Biology ; 552
ISBN: 9781603273169
Editor: Lundstrom, K.
Chiu, M.
Statement of
Responsibility: 
Tamara Cooper, Edward J. McMurchie and Wayne R. Leifert
Abstract: The [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.
Keywords: [35S]GTPγS
α2A-adrenergic receptor
G protein activation
DOI: 10.1007/978-1-60327-317-6_10
Appears in Collections:Aurora harvest
Molecular and Biomedical Science publications

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