Assay Method Information

Assay Name:  Protease Inhibition Assay
Description:  Inhibition of HIV protease was measured by assay of the cleavage of a fluorescent peptide substrate. The fluorescent product (2-aminobenzoyl-Ala-Thr-His-Gln-Val-Tyr) was separated from the fluorescent substrate by anion-exchange HPLC on a Mono-Q anion-exchange column (Pharmacia), and fluorescence was monitored at an excitation wavelength of 330 nm and an emission wavelength of 430 nm. Ki values for inhibitor binding were estimated at fixed concentrations of substrate and inhibitor from fractional activity measurements by use of the modified Michaelis-Menten equation.
Affinity data for this assay
 

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