West Nile Virus Protease, Fluorimetric 440, BioAssay(TM) Kit

Kit Size: 500 Assays (96-well)\n\nThis kit is optimized to detect West Nile Virus protease activity. It provides enough reagents to perform 500 assays in 96-well format.\n\nHigh Speed: The entire process only takes about half an hour.\n\nWest Nile virus (WNV) is from the family Flaviviridae3 found in both tropical and\ntemperate 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\nWNV contains a single-stranded, positive-sense RNA genome, which encodes three\nstructural proteins (capsid (C), membrane (M), envelope (E)), and seven non-structural proteins\n(NS1, NS2A, NS2B, NS3, NS4A, NS4B, NS5).5, 6 The West Nile viral NS3 protease is absolutely\nessential (along with viral-encoded cofactor NS2B) for post-translational processing of a viral\npolypeptide 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\n\nThe 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.\n\nKit Components:\nComponent A Microtiter Plate 1x500 wells Pyr-RTKR-AMC, WNV protease substrate, Ex/Em=354 nm/442nm 5mM DMSO solution, 1x250ul\nComponent B AMC, fluorescence reference standard, Ex/Em=354nm/442nm\n5 mM DMSO solution, 1x10ul\nComponent C Assay Buffer 2 x 50ml\nComponent D WNV Protease Inhibitor undeca-D-Arg-NH2 1 mM DMSO solution, 1x10ul\n\nMaterials Required (but not provided):\n

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SPECIFICATIONS

Catalog Number

W1018-69

Size

1Kit

Applications

ELISA

References

1. Shiryaev, SA. et al., Biochem J. 393, 503 (2006). 2. Chappell, KJ. et al., J. Biol. Chem. 281, 38448 (2006). 3. Hayes, CG. Ann. NY. Acad. Sci. 951, 25 (2001). 4. van der Meulen, KM et al., Arch. Virol. 150, 637 (2005). 5. Brinton, MA. Annu. Rev. Microbiol. 56, 371 (2002). 6. Lanciotti, RS. et al., Science 286, 2333 (1999). 7. Mueller, NH. et al., Int. J. Biochem. Cell Biol. 39, 606 (2007).