| Assay Method Information | |
| | 5-HT2C Receptor Binding Inhibition Assay |
| Description: | Binding Test of the Compound According to the Present Invention:In advance, 0.5 μL of the compound solution dissolved in DMSO was dispensed into a microplate, and the cell membrane and the hot ligand were diluted with Assay buffer, respectively. Then, the Assay buffer containing the diluted cell membrane was dispensed into a microplate at 50 μL/well. Then, the radioactive ligand solution was dispensed into a microplate at 50 μL/well, and the plate was sealed. Then, it was allowed to stand at 37° C. for 2 hours. During this period, 50 mmol/L Tris-HCl (pH 7.4) was dispensed into a GF/B UniFilter plate at 50 μL/well and allowed to stand at 4° C. for 1 hour or longer. After that, filtration was performed with Cell harvester (PerkinElmer). After the GF/B UniFilter plate was dried at room temperature, MicroScinti 20 was dispensed into the GF/B UniFilter plate at 50 μL/well, and the plate was sealed. The GF/B UniFilter plate was allowed to stand overnight at room temperature. The radioactivity of [3H]-Mesulergine bound to the 5-HT2A receptor was measured using Microbeta2 (PerkinElmer) at a measurement time of 1 min/well. The non-specific binding was calculated from the radioactivity of [3H]-Mesulergine in the presence of 500 μmol/L Serotonin HCl with the unlabeled ligand, and the total binding was calculated from the radioactivity of [3H]-Mesulergine in the absence of the compound according to the present invention (Vehicle). |
| Affinity data for this assay | |
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