HCV protease is identified as an important drug-screening target. The SensoLyte® 490 HCV Protease Assay Kit uses an optimized FRET peptide substrate for the continuous measurement of HCV NS3/4a activity. This FRET substrate is cleaved specifically by HCV NS3/4a protease thereby liberating the C-terminal peptide-fluorophore fragment from the proximity quenching effect of the dark quencher, resulting in a more than 10-fold increase in fluorescence. This assay can detect HCV NS3/4a protease activity at 1 pmole.
Component A: HCV NS3/4A protease substrate Ex/Em=340 nm/490 nm upon cleavage: 600 µL
Component B: EDANS, fluorescence reference standard: 100 µM DMSO solution, 20 µL
Component C: 2X Assay buffer: 50 mL
Component D: Stop solution: 30 mL
Component E: DTT: 1 M, 1 mL X 2 vials
Component :F Pep4AK, HCV NS3 protease cofactor: 300 µL, 600 µM
Storage & stability
Store all components at -20°C. Components C and D can be stored at room temperature for convenience. Protect Components A and B from light.