The SensoLyte® 620 HCV Assay Kit uses a new FRET peptide substrate that incorporates HiLyte Fluor™ TR (fluorophore) and QXL™ 610 (quencher) for a continuous measurement of enzyme activities. In the intact FRET peptide, the fluorescence of Hilyte Fluor™ TR is quenched by QXL™ 610. Upon cleavage of the FRET peptide by HCV NS3/4a protease, the fluorescence of Hilyte Fluor™ TR is recovered and can be continuously monitored at excitation/emission = 590 nm/620 nm. With near red emission wavelength, this HiLyte Fluor™ TR/QXL™ 610-based FRET peptide avoids the autofluorescence interference from most test compounds. This feature makes this FRET peptide an ideal substrate for screening HCV protease inhibitors since some of screening compounds have strong autofluorescence at shorter wavelength, making other FRET peptide substrates inappropriate. The assays are performed in a convenient 96-well or 384-well microplate format.
The kit contains: • HiLyte Fluor™ TR/QXL™ 610-based FRET peptide substrate (Ex/Em=590/620 nm upon cleavage) • Assay buffer • Fluorescence reference standard for calibration • A detailed protocol