Zika Virus Reporter Virus Particle (RVP) with Renilla Luciferase Reporter
Catalog Number:
X40H97
Description:
Reporter virus particles (RVPs) package the capsid-preMembrane-envelope (C-prM-E) gene construct of ZIKV (GenBank: KU321639.1, SPH2015 strain) and a Renilla luciferase reporter. As the wild typed Zika virus is handled under BSL 2 precautions, this Zika virus RVP can be handled using BSL-2 containment practices. Infection of cells with this RVP carrying Renilla luciferase reporter results in high level Renilla luciferase activity. See our titration result showing that the starting 2-fold diluted RVP-infected target cells (Vero) generates signal ~ 100,000-fold higher than uninfected control (cell alone as background in blue). We also evaluated the neutralizing activity of two published monoclonal antibodies (mAbs) – domain Ⅲ specific mAb ZV67 and mAb EDE1-C8 that binds to quaternary epitope. See figure
Applications:
- Anti-Zika virus neutralizing antibody screening at high throughput
- Anti-Zika virus drug screening at high throughput
- Zika virus vaccine efficacy evaluation at high throughput
- Zika virus pseudovirus transduction of target cells for viral entry, receptor recognition, cellular tropism and functional studies, such as ADCC or ADE analysis
Biosafety Level:
Handle it in biosafety cabinet in BSL-2. Contacted tips & tubes should be decontaminated by 10% disinfecting bleach
Organism:
Zika virus
Storage and shipping condition:
Shipping with dry ice. Require -80°C storage. Multiple freeze/Thaw cycles will reduce its sensitivity. Recommend only 1 cycle. Aliquot after the first thaw.
Size:
200 μl with recommended 1,000-fold dilution. We recommend to use it at a dilution fold where the signal of RVP infection is 1,000-fold higher than uninfected control (cell alone as background), although 100-1,000-fold high is acceptable. Can be used for 2,000 reactions (100 μl diluted virus) or 5,000 reactions if using reduced amount of RVP (40 μl diluted virus).
Use protocol:
The complete protocol is shipped with the product.
Briefly, incubate 100 μl diluted virus with 100 μl your sample in each well of 96-well plate with at least one duplicate for 30-60 min at 37°C. Then add 100 μl target cells (Vero or others expressing receptor). Read Renilla luciferase signal next day. For protocol using reduced amount, incubate 40 μl diluted virus with 10 μl your sample in each well of 96-well plate for 30-60 min at 37°C. Then add 25 μl target cells. Add 150ul fresh media next day and read signal on Day 3.
Note:
- Pseudoviruses are intended for Research Use Only
- A positive control mAb is free with pseudovirus purchase
