| Assay Method Information | |
| | [3H] LSD Binding Assay |
| Description: | For receptor binding assays, human 5-HT6 and 5-HT7 serotonin receptors expressed in CHO-K1 cells (PerkinElmer) were used. For 5-HT6 receptor binding, frozen membrane, 1 nM [3H]LSD (lysergic acid diethylamide; PerkinElmer), and appropriate concentrations of test compounds were added to 0.25 mL of 50 mM Tris-HCl (pH 7.4) buffer containing 0.1% ascorbic acid and 4 mM CaCl2. Nonspecific binding was determined using 10 μM methiothepin. The incubations were performed for 60 min at 27 °C, and these were terminated by rapid filtration through a Filtermat A glassfiber filter presoaked in 0.5% PEI. For the 5-HT7 receptor binding assay, cell membrane, 3 nM [3H]LSD, and appropriate concentrations of test compounds were added to 0.25 mL of 50 mM Tris-HCl (pH 7.4) buffer containing 10 mM MgSO4 and 0.5 mM EDTA. The mixture was incubated for 120 min at 27 °C, and the reaction was terminated by rapid filtration through a Filtermat A glass fiber filter presoaked in 0.3% PEI. Nonspecific binding was determined using 10 μM methiothepin. |
| Affinity data for this assay | |
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