CHEMBL3705175

Functional Assay: To measure D2 and D3 stimulation of mitogenesis (agonist assay) or D2 and D3 inhibition of quinpirole stimulation of mitogenesis (antagonist assay), CHOp-cells (human receptor) were seeded in a 96-well plate at a concentration of 5,000 cells/well. The cells were incubated at 37C. in alpha -MEM with 10% FBS, 0.05% penicillin-streptomycin, and 200 ug/mL of G418. After 48 hours, the cells were rinsed twice with serum-free alpha -MEM and incubated for 24 hours at 37 C. In the functional assay for agonism, the medium was removed and replaced with 90 ul of serum-free alpha -MEM and 10 ul of test compound in sterile water; in the antagonist assay, the test compound was diluted in sterile water plus 30 nM quinpirole. After another 24-hour incubation at 37 C., 0.25 uCi of [3H]thymidine was added to each well and the plates were further incubated for 2 hours at 37 C. The cells were then trypsinized, and the plates were filtered and counted as usual in the art.