Table 2Detailed protocol of DiscoveRx D2 β-arrestin secondary assay

SequenceParameterValueDescription
1Cells3 μL2,100 cells/well
2Time16–20 hrIncubate at 37 °C and 5% CO2
3Ligand1 μL1.5 μM dopamine in HBSS+10 mM HEPES+SMB
4Reagent23 nLCompound library, dopamine as control (in DMSO)
5Time90 minIncubate at 37 °C and 5% CO2
6Detection Reagent1.5 μL1:5:19 Galacton Star Substrate: Emerald II solution: PathHunter buffer
7Time60 minRoom temperature incubation
8Detector30 secLuminescent settings, ViewLux plate reader

From: Discovery, optimization, and characterization of a novel series of dopamine D2 versus D3 receptor selective antagonists

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