Insect GeneJuice^® Transfection Reagent

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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 Reagent is 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.

 

	Lane Sample Yield (µg) M Perfect Protein™ Markers 1 b-gal, Crude 2 b-gal, Eluate 137 3 Fluc, Crude 4 Fluc, Eluate 123 5 MAP kinase, Crude 6 MAP kinase, Eluate 91 7 GST MAP kinase fusion, Crude 8 GST MAP kinase fusion, Eluate 89 9 cdc2 kinase, Crude 10 cdc2 kinase, Eluate 63 11 GST cdc2 fusion, Crude 12 GST cdc2 fusion, Eluate 63
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.
For more information on Insect GeneJuice see TB359.