| Assay Method Information | |
| | 5-HT2A Receptor Binding Inhibition Test |
| Description: | Radioactive ligand: [3H]-Ketanserin with a final concentration of around Kd value calculated by the following method[1046]Non-specific ligand: Serotonin HCl with a final concentration of 500 μmol/LThe Kd value is calculated when the lot of cell membrane is changed. In advance, 0.5 μL of a 1 mmol/L compound for non-specific binding calculation dissolved in DMSO or DMSO is dispensed into a microplate, and the cell membrane is diluted with an Assay buffer. The radioactive ligand solution is serially diluted and the count is confirmed with a liquid scintillator. Assay buffer containing diluted cell membrane is dispensed into a microplate at 50 μL/well. Then, the radioactive ligand solution is dispensed into a microplate at 50 μL/well, and the plate is sealed. It is allowed to stand at room temperature (25° C.) for 1.5 hours. During this period, 50 mmol/L Tris-HCl (pH 7.4) is 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 is performed with Cell harvester (PerkinElmer). The radioactive ligand solution is dispensed into an empty well of the GF/B UniFilter plate at 10 μL/well. After the GF/B UniFilter plate is dried at room temperature, MicroScinti 20 is dispensed into the GF/B UniFilter plate at 50 μL/well to seal the plate. The GF/B UniFilter plate is allowed to stand overnight at room temperature. The radioactivity of [3H]-Ketanserin bound to the 5-HT2A receptor is measured using Microbeta2 (PerkinElmer) at a measurement time of 1 min/well. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |