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Het meten van G-eiwit-gekoppelde Receptor Signaling via Radio-gelabeld GTP Binding
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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
Chapters
- 00:05Title
- 00:37Expression of Recombinant HA-MOR1, Cell Fractionation, and Membrane Collection
- 03:55[35S]GTPγS Binding
- 06:05Membrane Filtration
- 06:59Liquid Scintillation Counting
- 07:57Results: Characterization of G-protein-coupled Receptors by Monitoring Radio-labled GTP Binding
- 09:01Conclusion
Guanosine trifosfaat (GTP) binding is een van de vroegste gebeurtenissen bij G-proteïne-koppelde receptor (GPCR) activatie. Dit protocol beschrijft hoe u specifieke GPCR-ligandinteracties farmacologisch kunt karakteriseren door de binding van het radioactief gemerkte GTP-analoog, [35S] guanosine-5'-O- (3-thio) trifosfaat ([35S] GTPγS) te monitoren in Reactie op een ligand van interesse.
