| Assay Method Information | |
| | [3H]-Gabapentin Binding Assay |
| Description: | A test compound (20 μL) diluted to 5 times of the final concentration with Binding buffer, [3H]-gabapentin diluted to 100 nmol/L with binding buffer [20 μL (final concentration 20 nmol/L)], and the rat cerebral cortex membrane fraction (60 μL; 12 μg membrane fraction) obtained in the (1) above were added to each well of a 96-well round-bottom plate. After being sufficiently mixed, these were allowed to react at room temperature for 1 hour. After the reaction, the reaction sample was suction filtered using a filter plate with 50 μL/well of 0.3 vol % polyethyleneimine added, and a cell harvester. The filter was then washed with ice-cooled Wash buffer (100 mmol/L NaCl, 0.1 w/v % BSA). After being washed, the filter plate was dried, and a scintillation cocktail (MicroScint-20, purchased from PerkinElmer) was added (50 μL/well), and the radioactivity on the filter was measured. The radioactivity obtained in the absence of the test compound was measured as total binding amount, and the radioactivity obtained by addition of an unlabeled gabapentin (final concentration 100 mmol/L) as a test compound was measured as non-specific binding amount. |
| Affinity data for this assay | |
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