Insect GeneJuice® Transfection Reagent

DNA Transfection Reagent for insect cells

Insect GeneJuice^® Transfection Reagent is a proprietary liposome formulation optimized for maximal transfection efficiency of insect cells including Sf9, S2, and HiFive^® cells. The reagent features extremely low toxicity to the cells and can be used for both transient and stable transfections in serum-containing or serum-free media, and for cotransfection of transfer plasmids with linearized virus DNA for the production of recombinant baculovirus. Insect GeneJuice is ideal for large-scale protein expression in suspension-culture transfections of Sf9 insect cells when using pIEx™ and pBiEx™ vectors, which contain the immediate early baculovirus promoter, IE1.

Insect GeneJuice Transfection Reagentis provided as a 2 mg/ml suspension in 20 mM MES, 150 mM NaCl, pH 6.2 buffer. One milliliter is sufficient for 10 transfections in 10-ml suspension-culture flasks or 100 transfections in 35-mm plates.

Insect GeneJuice Reagent

Target protein expression level and purification from transfected Sf9 cells
Sf9 cultures in 10-ml suspension cultures (1 x 10⁶ cell/ml) were transfected with 20 µg of the indicated plasmids using Insect GeneJuice Transfection Reagent. Total cell extracts were prepared 48 h later by the addition of Insect PopCulture™ Reagent (500 µl) followed by the addition of Benzonase^® Nuclease (5 µl). Samples were taken at this point to represent the total cell protein. Ni-NTA His•Bind^® Resin (50 µl per culture) was then added to the extracts. The 10-ml cultures were processed robotically using a MultiPROBE® II HT EX Liquid Handling Station (PerkinElmer). Target proteins purified from the 10-ml cultures were eluted in a volume of 150 µl. Crude and purified eluate fractions (10 µl each ) of the from each transfection were loaded in adjacent lanes of a 10–20% SDS polyacrylamide gel, which was stained with Coomassie™ blue. Purified protein yields were determined by BCA assay

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