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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 |
Published version: | http://dx.doi.org/10.1007/978-1-60327-317-6_10 |
Appears in Collections: | Aurora harvest Molecular and Biomedical Science publications |
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