West Nile Virus Envelope Protein ELISA Kit

Sample

Cell or tissue samples

Detection Method

sELISA

Intended Use

The West Nile Virus Envelope Protein ELISA Kit is an enzyme immunoassay designed to measure WNV E Protein from cell or tissue samples. It provides sufficient reagents for up to 96 tests in a 96-well plate including standard curve and unknown samples. Detection sensitivity is 31.3 ng/mL.

Contents of Kit

Box 1 (shipped at room temperature)
1. Anti-WNV E Protein Antibody Coated Plate: One 96-well strip plate (8 × 12).
2. Biotinylated Anti-WNV E Protein Antibody (1000×): One 10 μL vial.
3. Streptavidin-Enzyme Conjugate: One 20 μL vial.
4. Assay Diluent: One 50 mL bottle.
5. Lysis Solution: One 15 mL bottle containing 0.5% Tween 20 and 1% Triton X-100 in 10× PBS. The lysis solution was developed according to Mayo and Beckwith, 2002 and Colavita et al, 2017.
6. 10× Wash Buffer: One 100 mL bottle.
7. Substrate Solution: One 12 mL amber bottle.
8. Stop Solution: One 12 mL bottle.

Box 2 (shipped on blue ice packs)
1. WNV E Protein Standard: One 50 μL vial of 100 μg/mL WNV E Protein.

Storage

Upon receipt, aliquot and store the WNV E Protein Standard at -80°C and the Biotinylated Anti-WNV E Protein Antibody at -20°C. Avoid multiple freeze/thaw cycles. Store all other components at 4°C.

General Description

West Nile virus (WNV) is a single-stranded RNA virus that causes West Nile fever and is a member of the genus Flavivirus, which includes Zika virus, dengue virus, and yellow fever virus. WNV is transmitted mostly by mosquitoes, but the primary hosts of WNV are birds and the transmission cycle occurs from bird to mosquito to bird.
The WNV genome is a positive single stranded RNA encased by a nucleocapsid which is contained in a lipid bi-layered 50 nm envelope. A single polyprotein is expressed and cleaved by host and viral proteases into three structural (C, prM/M, and envelope, or E protein) and seven nonstructural (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5) proteins. The WNV E protein is responsible for cellular entry of WNV. The crystal structure of the WNV E protein shows that there are three domains: a βbarrel-shaped domain I, an elongated finger-like domain II, and a C-terminal immunoglobulin-like domain III8. Domain I is glycosylated at amino acid position 154, which is critical for WNV infection. The internal fusion peptide loop in domain II allows for the trimerization of the WNV E protein as well as initiation of virus entry into cells. Domain III controls binding of WNV to host cells.

Standard Curve

The following figures demonstrate typical results with the West Nile Virus Envelope Protein ELISA Kit. One should use the data below for reference only. This data should not be used to interpret actual results.

General Notes

Infectious Disease ELISA Kits