ViraPower™ HiPerform™ Lentiviral FastTiter™ Gateway® Expression Kit

Catalog number: K534000

Invitrogen™  Related applications: Cellular Imaging | Protein Assays and Analysis | Protein Expression

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Description

The ViraPower™ HiPerform™ Lentiviral FastTiter™ Gateway® Expression Kit includes all the components needed to generate lentivirus, including vector kit, 293FT cell line, and the support kit. This kit combines Invitrogen’s ViraPower™ HiPerform™ Lentiviral and Gateway® technologies to facilitate easy recombination-based cloning and lentiviral-based, high-level expression of a target gene in dividing and non-dividing mammalian cells. The pLenti7.3⁄V5-DEST™ vector is equipped with two key genetic elements, making it a HiPerform™ vector: the Woodchuck Posttranscriptional Regulatory Element (WPRE) and the central Polypurine Tract (cPPT) sequence from the HIV-1 integrase gene to produce at least 4-fold increase in protein expression compared to vectors lacking these elements. In addition, the vector allows for an accurate determination of titer of functional lentivirus in just two days using Emerald Green Fluorescent Protein (EmGFP).

Advantages
• Transient expression
• Short-term experiments
• High-throughput screening
• 2 day titer of functional virus using EmGFP
• Flexible and versatile Gateway® recombination cloning technology

Key Features
• WPRE (Woodchuck Posttranscriptional Regulatory Element) from the woodchuck hepatitis virus, allows increased transgene expression and cPPT (Polypurine Tract) from the HIV-1 integrase gene, increases the copy number of lentivirus integrating into the host genome
• Human cytomegalovirus (CMV) immediate early promoter to control high-level expression of the gene of interest
• SV40 promoter driving expression of EmGFP
• Emerald Green Fluorescent Protein (EmGFP) allows easy determination of Lentiviral titers by flow cytometry and monitor transduced cells.

Kit includes
• pLenti7.3⁄V5-DEST™ Gateway® Vector Kit (V534-06)
• 293FT Cell Line (R700-07)
• ViraPower™ Lentiviral Support Kit (ViraPower™ Lentiviral Packaging Mix and Lipofectamine® 2000)

For research use only. Not intended for any therapeutic or diagnostic use.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Promoter: CMV
Key Function: Viral Expression, Reporter Assays
Protein Tag or Fusion: V5 Epitope Tag
Reporter: GFP (EmGFP)
Delivery Method: Lentiviral
Cloning Method: Gateway®
Selection Agent (Eukaryotic): None
Vector Type: pLenti
Product Size: 1 kit
Special Elements: WPRE, cPPT
Constitutive or Inducible System: Constitutive

Contents & storage

pLenti7.3⁄V5-DEST™ Gateway® Vector Kit contains pLenti7.3⁄V5-DEST™ Gateway® Vector, 40 µl at 150 ng⁄µl and pLenti7.3⁄V5-GW⁄lacZ Control Vector, 20 µl at 0.5 µg⁄µl. Store at -20°C.

ViraPower™ Lentiviral Support Kit contains enough reagents for 20 transfections in 10 cm tissue culture dishes: 195 µg lyophilized mix Packaging Mix. Store at-20°C; 0.75 ml Lipofectamine® 2000. Store at +4°C.

One Shot® Stbl3™ chemically competent E. coli box contains transformation reagents including 20 single-use 50 µl aliquots of One Shot® Competent Cells, S.O.C. medium, and pUC19 supercoiled plasmid control. Store at -80°C.

293FT cell line: 3 x 106 frozen cells in 1 ml of Freezing Medium. Store in liquid nitrogen.

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Manuals & protocols

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