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Assay Method Information

Assay Name:  Measurement of GPCR-mediated thallium flux through GIRK channels: Dose-Response Testing
Description:  Assay Provider: Colleen Niswender Assay Provider Affliation: Vanderbilt University Grant Title: Measurement of GPCR-mediated thallium flux through GIRK channels Grant Number: 1 R03 MH076398-01 The aim of this work was to use high throughput screening of a small molecule library to identify compounds that interact with the alpha2C adrenergic receptor. The assay utilized thallium influx through G-protein Inwardly Rectifying K+ (GIRK) channels as a measure of alpha2C activation. Compounds were tested at 10uM final concentration against Human Embryonic Kidney (HEK) 293 cells stably expressing endogenous alpha2C and cDNAs for GIRK 1 and GIRK 2. The Hamamatsu FDSS 6000 plate reader was used to collect kinetic fluorescence intensities during treatment with the test compound. The alpha2C agonist UK-14304 (Tocris) was the positive control, and DMSO, the compound vehicle, was used as the negative control.
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Last update November 1, 2007
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