| Assay Method Information | |
| | Radioligand-Receptor Binding Assay |
| Description: | Dopamine D3 Receptor: Experimental buffer: 50 mM Tris-HCl pH 7.4, 10 mm MgCl2; washing liquor: 50 mM Tris-HCl pH 7.4, stored at 4° C.; 0.5% PEI solution: 0.5 g PEI dissolved in 100 mL ddH2O, 4° C. storage of spare.5 μL of the test compounds (0.005 nM to 100 nM, 10 concentrations in total) and 100 μL of buffer were added to a 96-well assay plate. 1 μL of cell membrane and 300 μL of buffer were added to each well, and the plate was shaken at 600 rpm for 5 minutes. 100 μL of a mixed solution of buffer and [3H]-methylspiperone (final concentration of 0.5 nM) was added to the reaction system per well, and the plate was shaken at 600 rpm for 5 minutes and incubated at 27° C. for 30 min. The UNIFILTER-96 GF/B filter plate pre-incubated with 0.5% PEI for 1 h was washed twice with the buffer (1 mL/well). The cell membrane suspension was added to the UNIFILTER-96 GF/B filter plate, washed 4 times, and incubated at 55° C. for 10 min. 40 μL of ULTIMA GOLD was added to each well, and liquid scintillation counting was carried out. |
| Affinity data for this assay | |
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