| Assay Method Information | |
| | Dopamine D2L Receptor Binding Assay |
| Description: | The human D2L receptor binding activity of [3H]-spiperone was determined as follows. A solution containing 50 μL of [3H]-spiperone (the final concentration: 0.5 nmol/L), 2 μL of a solution of a test compound in DMSO or a solvent (DMSO), and 148 μL of human D2L receptor-expressed CHO cell membrane sample was reacted in a 50 mmol/L Tris-HCl (pH=7.6) buffer, and then was let stand at room temperature for 60 minutes. Then, the resultant was added promptly to a glass fiber filter plate (Multiscreen FB, Millipore) coated with 0.3% polyethyleneimine (PEI) and filtered under reduced pressure. The filtered substance on the glass fiber filter was washed with 200 μL of ice-cooled 50 mmol/L Tris-HCl (pH=7.6) twice and repeated to be filtered under reduced pressure, followed by transfer to a vial containing 2 mL of Ecoscint A (National Diagnostics). The radioactivity of the filtered substance remained on the glass fiber filter was measured by liquid scintillation counter. The radioactivity value measured by liquid scintillation counter was deemed to be a receptor binding activity, and the binding inhibition was calculated from the total binding (TB), non-specific binding (NSB), and specific binding (SB) of a test drug according to the following equations. |
| Affinity data for this assay | |
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