Kit Size: 500 Assays (96-well)
This kit is optimized to detect West Nile Virus protease activity. It provides enough reagents to perform 500 assays in 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 The 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 440 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 a fluorogenic peptide described in the literature.1 This peptide, Pyr-RTKR-AMC generates the AMC (7-amino-4-methylcoumarin) fluorophore upon NS3 protease cleavage. AMC has bright blue fluorescence that can be detected with excitation at 354nm and emission at 442nm.
Component A Microtiter Plate 1x500 wells Pyr-RTKR-AMC, WNV protease substrate, Ex/Em=354 nm/442nm 5mM DMSO solution, 1x250ul
Component B AMC, fluorescence reference standard, Ex/Em=354nm/442nm
5 mM DMSO solution, 1x10ul
Component C Assay Buffer 2 x 50ml
Component D WNV Protease Inhibitor undeca-D-Arg-NH2 1 mM DMSO solution, 1x10ul
Materials Required (but not provided):