| Assay Method Information | |
| | [3H]Epibatidine Binding Assay |
| Description: | Adult male rat cerebral cortices (Pelfreeze Biological, Rogers, Ark.) were homogenized in 39 volumes of ice-cold 50 mM Tris buffer (pH 7.4 at 4° C.) containing 120 mM NaCl, 5 mM KCl, 2 mM CaCl2, and 1 mM MgCl2 and sedimented at 37,000 g for 10 min at 4° C. The supernatant was discarded, the pellet resuspended in the original volume of buffer, and the wash procedure repeated twice more. After the last centrifugation, the pellet was resuspended in 1/10 its original homogenization volume and stored at −80° C. until needed. In a final volume of 0.5 mL, each assay tube contained 3 mg wet weight male rat cerebral cortex homogenate (added last), 0.5 nM [3H]epibatidine (NEN Life Science Products, Wilmington, Del.) and one of 10-12 different concentrations of test compound dissolved in buffer (pH 7.4 at room temperature) containing 10% DMSO resulting in a final DMSO concentration of 1%. Total and nonspecific bindings were determined in the presence of vehicle and 300 μM (−)-ni |
| Affinity data for this assay | |
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