| Assay Method Information | |
| | Affinity of the Compounds of the Present Invention for Dopamine D2 Receptors |
| Description: | The affinity of the compounds of the present invention for the dopamine D2 receptors was determined by the method of radioligand competition experiment. In the first step, a cell membrane component containing specific dopamine D2 receptors was prepared. A10 cm dish was used for transfection with 10 ng of the dopamine D2 receptors and 40 μL of PEI. After 48 hours, the 10 cm dish was taken out from the cell culture room and the cultured cells had expressed the dopamine D2 receptors. A vacuum pump was used to suck off the culture medium, 3 mL of lysis buffer was added to each well, and the cells were placed in a 4° C. freezer for 10 minutes. After the cells were detached, the cells were transferred to a 15 mL centrifuge tube and centrifuged at 1500 rpm for 5 minutes at 4° C., and the supernatant was discarded. The cell pellet was transferred to a tissue homogenizer, and 3 mL of lysis buffer was added and fully ground until the cells were broken. Then, cell suspension was equally divided into several EP tubes, centrifuged at 4° C. and 12000 rpm for 5 min, and the supernatant was discarded. The precipitate was the cell membrane component containing the dopamine D2 receptors. In the second step, a ligand-receptor binding experiment was performed on HEK-293T membrane component transiently expressing the dopamine D2 receptors. |
| Affinity data for this assay | |
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