| Assay Method Information | |
| | Binding Assay for gamma2-Containing GABAA Subtypes |
| Description: | Radioligand binding assays were carried out in a volume of 200 μL (96-well plates) which contained 100 μL of cell membranes, [3H]Flumazenil at a concentration of 1 nM and the test compound in the range of [0.1·10−3−10]×10−6 M. Nonspecific binding was defined by 10−5 M Diazepam and typically represented less than 5% of the total binding. Assays were incubated to equilibrium for 1 hour at 4° C. and harvested onto GF/C uni-filters (Packard) by filtration using a Packard harvester and washing with ice-cold wash buffer (50 mM Tris; pH 7.5). After anhydrousing, filter-retained radioactivity was detected by liquid scintillation counting. Ki values were calculated using Excel-Fit (Microsoft) and are the means of two determinations. |
| Affinity data for this assay | |
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