Assay Method Information

Assay Name:  Receptor Binding Assay
Description:  The affinity of the test compounds for the GABA gated Cl-channel (GABA.A receptor complex) in the rat cerebral cortex was determined in a radioligand binding assay (Lewin et al,. 1989). Membrane homogenates of cerebral cortex are incubated for 120 min at 22° C. with 3 nM [35S]TBPS (-t-butylbicyclophosphoorothionate) in the presence and absence of the test compound.Following incubation, the samples are filtered rapidly under vacuum through glass fiber filters and rinsed several times with ice-cold buffer using a 96-sample cell harvester. The filters are dried then counted for radioactivity in a scintilliation counter using a scintillation cocktail (Microscint 0, Packard). Each compound was tested at several concentrations to calculate the IC50 values of the affinity to the GABA.A receptor complex in cerebral cortex of rats.
Affinity data for this assay
 

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