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Biochemistry

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JoVE Journal Biochemistry
Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding
 

Measuring G-protein-coupled Receptor Signaling via Radio-labeled GTP Binding

Article DOI: 10.3791/55561-v • 10:13 min • June 9th, 2017
June 9th, 2017 •
*1, *1, 1, 1,2, 1,3,4
1The Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, 2Department of Neurology and neurosurgery, Johns Hopkins University School of Medicine, 3Departments of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, 4Department of Psychiatry and Behavioral Sciences, Johns Hopkins University School of Medicine
* These authors contributed equally

챕터

요약
Automatic Translation

Guanosine triphosphate (GTP) binding is one of the earliest events in G-Protein-Coupled Receptor (GPCR) activation. This protocol describes how to pharmacologically characterize specific GPCR-ligand interactions by monitoring the binding of the radio-labeled GTP analog, [35S]guanosine-5'-O-(3-thio)triphosphate ([35S]GTPγS), in response to a ligand of interest.

Tags

G-protein-coupled Receptor Signaling Radio-labeled GTP Binding Pharmacology Biochemistry Cellular Membranes Functional GTP Binding Assay Plate Cells Transfection Incubation Rinse Cells Buffer 1 Cell Scraper Micro Centrifuge Tube Lysates Thaw
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