| Assay Method Information | |
| | Receptor Binding Assay |
| Description: | Receptor binding assays were performed in 96-well format in deep-well plates. For each 96-well plate one ampule of membrane homogenate was thawed and diluted in binding buffer (50 mM Tris pH=7.4, 100 mM KCl) and 200 uL was deipensed into each well. Radioligand [3H]Ro151788 (Perkin Elmer: NET757250UC) was prepared in binding buffer and added to each well in 50 uL volumne to give final concentration of 0.5 nM. Test compounds in suitable concentrations(s) were added in additional 50 uL. The final asssay volume was 300 uL. Incubation was carried out for 60 minutes at 4C. for non-specific binding 10 uM unlaved diazepam was used. After incubation samples were filtered over UniFilter GF/BT using Filtermate Harvester (Perkin elmer)and washed was 5x1 mL binding buffer. The plate was dried at 40C for an hour and 40 uL Microscint (Perkin Elmer) sintillation cocktail was added to each well. The plate was read in Microbeta (Perkin Elmer). The specific radioligand binding (SB) was defined as the difference between total binding (Tot) and the non-specific binding (NSB). |
| Affinity data for this assay | |
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