Assay Method Information

Assay Name:  [3H]GABA Filtration Binding Assay (Ki) and FLIPR Assay for Agonism (EC50)
Description:  Inhibition of [3H]GABA binding at GABAB receptor sites in rat brain synaptic membranes by test compounds was measured using a filtration binding assay. Displacement curves to determine IC50 values were constructed by fitting the 4-parameter logistic equation to the data. Kd for GABA was determined on each preparation by homologous competition and used to calculate Ki values from IC50 determinations using the Cheng-Prusoff equation. The average Kd for GABA was determined to 110 +/- 21 nM. The EC50/IC50 values were obtained from measurement of GABAB receptor dependent release of intracellular calcium in Fluorescence Imaging Plate Reader (FLIPR) assay using CHO cells transfected with GABAB(1a)-Gqi5 and GABAB(2).
Affinity data for this assay

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