| Assay Method Information | |
| | Dopamine D2 Receptor Activity Assay |
| Description: | D2 receptor/G protein α16a co-transfected HEK293 cells (from the Shanghai Institute of Materia Medica, Chinese Academy of Sciences) were seeded in 96-wellplates and incubated overnight at 37 °C. The media were then removed, and 40 μL of 10 μM Fluo-4/AM (a calcium fluorescent probe) (Invitrogen Corporation, Carlsbad, CA, USA) was added. The plates were then incubated for 40 min at 37 °C. After the supernatant was discarded and the cells were washed with calcium buffer, 50 μL of calcium buffer containing tested compounds at the dosage range from 0.01 nM to 10 μM was added, and the plates were incubated for 10 min. The fluorescence values at an emission wavelength of 525 nm and an excitation wavelength of 485 nm were detected by the FlexStation II microplate reader (Molecular Devices, San Francisco, CA, USA), starting 15 s after 25 μL of calcium buffer containing 50 nM dopamine (an agonist of the dopamine D2 receptor) (Sigma-Aldrich, Saint Louis, MO,USA) was automatically added by the instrument. |
| Affinity data for this assay | |
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