Assay Method Information

Assay Name:  Receptor Activity Assay
Description:  The compounds are tested on HEK-293 (Human Embryonic Kidney) cells stably expressing the subunit α5 of the human GABAA receptor, and also the subunits beta2 (short) and gamma2 (long). The cells are maintained in the presence of a selection of three antibiotics, neomycin, zeocin and puromycin in a Dulbecco medium (DMEM) containing 10% (v/v) foetal bovine serum. On the day before the experiment, the cells are transferred to 96-well plates (in a density of 50,000 cells/well). The cells are then preincubated for 40 minutes with the compounds under test and are treated with GABA. The membrane potential is monitored using a blue FMP marker (Molecular Devices), following the instructions of the manufacturer. The responses are recorded for 120 seconds on a FlexStation3 plate reader (Molecular Devices, USA).
Affinity data for this assay
 

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