| Assay Method Information | |
| | Screening for 5-HT2A Inverse Agonistic Activity |
| Description: | NIH3T3-5HT2AR cells in the logarithmic growth phase were seeded into a white-wall clear-bottom 96-well plate at a density of 1000 cells per well, and cultured overnight in a 37 C., 5% CO2 incubator. The next day, a compound to be tested was added to the cells, wherein the compound to be tested was subjected to 3.16-fold gradient dilution with PBS to obtain 9 concentrations, with the highest concentration of 10 uM, and double replicate wells were set up for each concentration; and PBS was used as a negative control group, and pimavanserin with the same concentration was used as a positive control group. After the addition, the cells were cultured in a 37 C., 5% CO2 incubator for another 120 h. On the sixth day, a Bright-Glo Luciferase reagent with a volume equal to that of cell sap was added to the cells, and the cells were incubated at room temperature in the dark for 20 min. The plate was shaken every 5 min with a plate shaker, the luminescent intensity was detected by a microplate reader, and the cell inhibition rate was calculated. The data was processed with GraphPad Prism 7.0 to obtain a cell inhibition rate curve, and the IC50 was calculated. |
| Affinity data for this assay | |
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