VSV Pseudovirus_ Sudan Ebola GP with luciferase reporter

Catalog Number:

U95M81

Description:

This pseudotyped virus uses recombinant vesicular stomatitis virus (rVSV) to carry GP protein of Sudan Ebolavirus strain (GenBank: KT878488.1). As pseudovirus infectivity of rVSV without its original G is restricted to a single round of replication, the pseudotypes can be handled using BSL-2 containment practices. Infection of cells with this pseudotyped virus carrying luciferase reporter results in high level luciferase activity. See our titration result showing that the starting 2-fold diluted pseudovirus generates signal over 100,000-fold higher than uninfected control (cell alone as background in blue). We also evaluated the neutralizing activity of one published mAb called CA45 by using this pseudotyped virus with a HIV control mAb, see results.

Applications:

• Anti-Ebolavirus neutralizing antibody screening at high throughput
• Anti-Ebolavirus drug screening at high throughput
• Ebolavirus vaccine efficacy evaluation at high throughput
• Ebolavirus pseudovirus transduction of target cells for viral entry, receptor recognition, cellular tropism and functional studies, such as ADCC analysis

Biosafety Level:

Handle it in biosafety cabinet in BSL-2. Contacted tips & tubes should be decontaminated by 10% disinfecting bleach

Organism:

Ebola virus -Related filovirus

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 100-fold dilution. We recommend to use it at a dilution fold where the signal of pseudovirus 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 200 reactions (100 μl diluted virus) or 500 reactions if using reduced amount of pseudovirus (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 for 30-60 min at 37°C. Then add 100 μl target cells (Vero). Read 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