A subscription to JoVE is required to view this content. Sign in or start your free trial.
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
Guanosintrifosfat (GTP) binding er en av de tidligste hendelsene i G-protein-koblet reseptor (GPCR) aktivering. Denne protokollen beskriver hvordan man farmakologisk karakteriserer spesifikke GPCR-ligand-interaksjoner ved å overvåke bindingen av den radiomerkede GTP-analogen, [35S] guanosin-5'-O- (3-tio) trifosfat ([35S] GTPγS), i Svar på en ligand av interesse.
