Assay Method Information

Assay Name:  Determination of Inhibition Concentration Values (IC50)
Description:  The inhibition concentration values of GCPII were determined using the radioenzymatic assay with [3H] NAAG (radiolabeled on the terminal glutamate). The enzyme was preincubated in the presence of increasing concentrations of inhibitors in reaction buffer. The reaction was initiated by adding the substrate to the reaction mixtures. After 20 min, the reaction was terminated, and the glutamate was separated from the reaction mixture by ion-exchange chromatography and quantified by liquid scintillation. Duplicate reactions were carried out for each experimental point. The IC50 values were calculated from plots of Vi/V0 (ratio of individual reaction rates to the rate of an uninhibited reaction) versus inhibitor concentration using the GraFit program (version 5.0.4, Erithacus Software Limited).
Affinity data for this assay
 

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