| Assay Method Information | |
| | Measurement of beta-arrestin Recruitment to Human Dopamine D3 Receptors |
| Description: | PathHunter CHO-K1 cells expressing tagged human D3 receptors and tagged β-arrestin-2 (Eurofins DiscoverX, Fremont, CA, USA) were seeded into 96-well white-walled clear bottom tissue culture plates in 90 μL AssayComplete™ Cell Plating 2 (CP2) reagent (Eurofins DiscoverX) at a density of 25,000 cells/well and incubated overnight in humidified atmosphere with 5% CO2 at 37° C. Twenty to twenty-four hours later, 20 μL of test compounds or vehicle in CP2 reagent containing 2.2% DMSO was added to the cells and the cells were incubated for 90 minutes at 37° C. Following incubation, 55 μL PathHunter® Detection Reagent (Eurofins DiscoverX) was added per well and the plates were incubated for 60 minutes at 25° C. followed by luminescence detection using a PHERAstar FS multimode plate reader (BMG Labtech, Ortenberg, Germany). |
| Affinity data for this assay | |
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