| Assay Method Information | |
| | Test for 5-HT2A Antagonistic Activity |
| Description: | CHO-K1/5-HT2A cells in the logarithmic growth phase were seeded into a 384-well plate at a density of 10000 cells per well, and cultured in a 37° C., 5% CO2 incubator for 16-24 h. After that, centrifugation was performed to remove the culture medium in the cell culture plate. Dye was added, and the cells were incubated in a 37° C., 5% CO2 incubator for another 1 h. The cell culture plate was placed on the FLIPR. A compound to be tested was added, and the Ca2+ signal was detected, wherein the compound to be tested was subjected to 3-fold gradient dilution with PBS to obtain 10 concentrations, with the highest concentration of 10 uM, and double replicate wells were set up. 100 nM a-methyl-5-HT was added 15 min later, and the Ca2+ signal was detected again, wherein the value obtained by only adding 100 nM a-methyl-5-HT was taken as the maximum signal. In order to determine the test stability, risperidone was used as a positive control. The data were 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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