[³⁵S]GTPγS binding in G protein-coupled receptor assays

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dc.contributor.authorCooper, T.
dc.contributor.authorMcMurchie, E.
dc.contributor.authorLeifert, W.
dc.contributor.editorLundstrom, K.
dc.contributor.editorChiu, M.
dc.date.issued2009
dc.description.abstractThe [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.
dc.description.statementofresponsibilityTamara Cooper, Edward J. McMurchie and Wayne R. Leifert
dc.identifier.citationMethods in Molecular Biology, 2009; 552:143-151
dc.identifier.doi10.1007/978-1-60327-317-6_10
dc.identifier.isbn9781603273169
dc.identifier.issn1064-3745
dc.identifier.issn1940-6029
dc.identifier.urihttp://hdl.handle.net/2440/57213
dc.language.isoen
dc.publisherHumana Press
dc.publisher.placeUnited States
dc.relation.ispartofseriesMethods in Molecular Biology ; 552
dc.rightsCopyright 2009 Springer
dc.source.urihttps://doi.org/10.1007/978-1-60327-317-6_10
dc.subject[35S]GTPγS
dc.subjectα2A-adrenergic receptor
dc.subjectG protein activation
dc.title[³⁵S]GTPγS binding in G protein-coupled receptor assays
dc.title.alternative[(35)S]GTPgammaS binding in G protein-coupled receptor assays
dc.typeJournal article
pubs.publication-statusPublished

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