This kit is optimized to detect West Nile Virus protease activity. It provides enough reagents to perform 100 assays in a 96-well format.
High Speed: The entire process only takes about half an hour.
West Nile virus (WNV) is from the family Flaviviridae3 found in both tropical and
temperate regions. The main route of human infection is through the bite of an infected mosquito Infection causes severe neurological disease and fatalities in both human and animal hosts. There is currently no effective vaccine or antiviral drug to protect against WNV infection.4
WNV contains a single-stranded, positive-sense RNA genome, which encodes three
structural proteins [capsid (C), membrane (M), envelope (E)], and seven non-structural proteins
(NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5).5,6 West Nile viral NS3 protease is absolutely
essential (along with viral-encoded cofactor NS2B) for post-translational processing of a viral
polypeptide precursor in infected host cells. This polypeptide provides the structural and functional viral proteins, and inhibition of its processing could represent a potential treatment for viral infections. Thus, this protease has become one of the key targets for developing anti-WNV drugs.1,7
The 570 West Nile Virus Protease Assay Kit provides a convenient,
homogeneous assay for high throughput screening of West Nile Virus protease NS3 inhibitors. It
allows for continuous quantification of protease activity using QXL(TM)570/5-TAMRA FRET
peptide. In the FRET peptide, the fluorescence of 5-TAMRA is quenched by QXL(TM)570. Upon
cleavage into two separate fragments by WNV protease, the fluorescence of 5-TAMRA is
recovered, and can be monitored at excitation/emission = 540 nm/575 nm. The long wavelength fluorescence of 5-TAMRA is less interfered by the autofluorescence of cell components and test compounds.
Component A Microtiter Plate 1x100 wells QXLTM570/5-TAMRA, WNV protease substrate, Ex/Em=540nm/575nm 1.5mM DMSO solution, 1x60ul
Component B 5-TAMRA, fluorescence reference standard, Ex/Em=540nm/575nm 750uM DMSO solution, 1x10ul
Component C Assay Buffer 1x20ml
Component D WNV Protease Inhibitor undeca-D-Arg- NH2 1mM DMSO solution, 1 1x0ul
Materials Required (but not provided):