Dengue Virus Reporter Virus Particle (RVP) with Renilla Luciferase Reporter (Type 2)

Catalog Number:

X57D33

Description:

Reporter virus particles (RVPs) package the capsid-preMembrane-envelope (C-prM-E) gene construct of Dengue virus (DENV) (GenBank: MK506263.1, Serotype 2 strain) and a Renilla luciferase reporter. As the wild typed Dengue virus is handled under BSL 2 precautions, this Dengue 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 (Huh-7) generates signal ~ 1,000-fold higher than uninfected control (cell alone as background in blue). We also evaluated the neutralizing activity of monoclonal antibody (mAb) – domain Ⅲ specific RV25 (Cat.No. K34Q76). See figure

Applications:

• Anti-Dengue virus neutralizing antibody screening at high throughput
• Anti-Dengue virus drug screening at high throughput
• Dengue virus vaccine efficacy evaluation at high throughput
• Dengue 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:

Dengue virus (Serotype 2)

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 10-fold dilution. We recommend to use it at a dilution fold where the signal of RVP infection is >100-fold higher than uninfected control (cell alone as background). Can be used for 20 reactions (100 μl diluted virus) or 50 reactions if using reduced amount of RVP (40 μl diluted virus).

Use protocol:

The complete protocol is shipped with the product. We strongly suggest that you use small aliquot to test your own systems and use our data as reference.

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 (Huh-7, 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 included with pseudovirus purchase