| Assay Method Information | |
| | GPR40 agonistic activity |
| Description: | hGPR40-CHO stably transfected cells were seeded into a 96-well plate at a density of 3×104/well, and incubated overnight in a 37° C., 5% CO2 cell incubator; the medium was discarded, 100 μl of HBSS was added to each well, and 100 μl of Fluo-4 dye solution containing Probenecid was added and incubated at 37° C. for 90 min. After the incubation, the Fluo-4 dye solution was sucked out, 100 μl of HBSS buffer was added, and the dye was washed away; 100 μl of HBSS containing Probenecid was added to each well, and incubated at 37° C. for 10 min; different concentrations of drugs were added to each well of the 96-well plate and FLIPR (Molecular Devices) was used for readings according to the parameter setting table. The experimental results were analyzed. Agonistic activity=(compound well fluorescence value−blank control well fluorescence value)/(linoleic acid well fluorescence value−blank control well fluorescence value)×100%, finally compounds with better agonistic activity were selected to determine their EC50 at 100 nM. |
| Affinity data for this assay | |
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