| Assay Method Information | |
| | Ca2+ Flux Assay |
| Description: | G ±15 HEK293 cells stably expressing rat mGlu2 were plated in black-walled, clear-bottomed, poly-D-lysine coated 384-well plates in 20 L of assay medium (DMEM containing 10% dialyzed FBS, 20 mM HEPES, and 1 mM sodium pyruvate) at a density of 12K cells/well. The cells were grown overnight at 37 ° C. in the presence of 5% CO2. The next day, medium was removed and the cells incubated with 20 L of 2.3 M Fluo-4, AM prepared as a 2.3 mM stock in DMSO and mixed in a 1:1 ratio with 10% (w/v) pluronic acid F-127 and diluted in assay buffer (Hank's balanced salt solution, 20 mM HEPES, and 2.5 mM probenecid) for 45 minutes at 37 ° C. Dye was removed, 20 L of assay buffer was added, and the plate was incubated for 5 minutes at room temperature. |
| Affinity data for this assay | |
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