In 2001, Novagen introduced GeneJuice^® Transfection Reagent for DNA delivery into mammalian cells. GeneJuice, a simple to use reagent, combines high transfection efficiency with exceptionally low cytotoxicity eliminating much of the optimization required by cationic liposome-based transfection reagents. This reagent has been used to successfully transfect many cell types including primary cells. Now, we are pleased to offer three additions to our transfection reagent or "juice" family: Insect GeneJuice™ Transfection Reagent (for insect cell transfections), RiboJuice™ siRNA Transfection Reagent, and ProteoJuice™ Protein Transfection Reagent. Each of our new transfection reagents carries on the tradition of GeneJuice by also being simple to use and highly efficient. Whether you are transfecting DNA, siRNA, or protein…Novagen has the juice for you!

GeneJuice^® Transfection Reagent is a proprietary formulation optimized for maximal transfection efficiency, ease of use, and minimal cytotoxicity. This transfection reagent is a superior alternative to newer lipid-based reagents, as well as traditional techniques including calcium phosphate coprecipitation, electroporation, microinjection, biolistic particle delivery, and complex formation with DEAE-dextran.

Whereas many available transfection reagents are based on cationic lipid formulations, GeneJuice is composed of a nontoxic cellular protein and a small amount of a novel polyamine. The unique chemistry provides several advantages over lipid-based transfection, including:

• Highly efficient DNA transfer for both stable and transient transfections
• Minimal cellular toxicity
• Compatibility with both serum-containing and serum-free media
• Simple protocol - no need for media changes
• Ideal for high throughput (HT) transfection in a multi-well plate

GeneJuice Transfection Reagent provides excellent performance in both stable and transient transfection of mammalian cells and is ideal for use with Novagen's pMLuc™, pTK-neo, pTriEx™, and pTandem™-1, reporter, drug slection, and expression vectors.

The 1 ml size provides enough reagent to perform up to 500 transfections in standard 35 mm plates. The reagent is also available in an introductory 0.3 ml size. GeneJuice is supplied as a ready-to-use sterile solution.

See the list of cell lines and primary cells successfully transfected with GeneJuice Transfection Reagent!

Insect GeneJuice^® Transfection Reagent is a proprietary liposome formulation optimized for maximal transfection efficiency of insect cells (SF9, S2, and HiFive^® cell transfections). 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.

M- Perfect Protein™ Markers bgal - Crude bgal - Elute Yield =137µg Fluc - Crude Fluc- Elute Yield =123µg MAP kinase - Crude MAP kinase - Elute Yield =91µg GST MAP kinase fusion- Crude GST MAP kinase fusion - Elute Yield =89µg cdc2 kinase - Crude cdc2 kinase - Elute Yield =63µg GST cdc2 fusion - Crude GST cdc2 fusion - Elute Yield =63µg

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 1-ml cultures were processed manually and the 10-ml cultures were processed robotically using a MultiPROBE® II HT EX Liquid Handling Station (PerkinElmer). Target protein purified from the 1- and 10-ml cultures was eluted in a volume of 100 µl and 150 µl, respectively. Ten microliters of the crude and purified fractions 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.

RiboJuice™ siRNA Transfection Reagent efficiently delivers small interfering RNA (siRNA) into a wide range of mammalian cell lines for targeted gene suppression. When transfected into the cell, the siRNA directs cleavage of the target message resulting in suppression of protein expression without affecting non-target genes. RiboJuice is effective for siRNA delivery in a variety of cell lines with multiple siRNAs (see data below). The ability to target specific genes for gene silencing is a powerful tool that researchers can now perform more easily, using RiboJuice siRNA Transfection Reagent.

Concentration dependence and specificity of siRNA(b-gal)-mediated suppression CHO-K1 and HEK-293 cells plated at 50,000 cells per well were transfected after 24 h with two mixtures. Mixture one contained 1 µl GeneJuice™ Transfection Reagent, 0.25 µg pTriEx™-2(bgal), and 0.025 µg pTriEx-2(Fluc). Mixture two contained 3 µl RiboJuice™ and either 0.1, 0.25, 0.5, 1, 2.5, 5, 10, or 25 nM (final concentration) of the indicated siRNAs. As a control, 3 µl RiboJuice was also used without any siRNA. Total volume per well was 300 µl. Cells were lysed with Reportasol™ Extraction Buffer 24 h after transfection and assayed for reporter activity. All assays were performed in triplicate and variation is expressed as standard error of the mean.

Suppression of b-galactosidase expression in various cell lines The indicated cell lines were plated at 50,000 cells per well and transfected after 24 h with two mixtures. Mixture one contained 1 µl GeneJuice Transfection Reagent, 0.25 µg pTriEx-2(b-gal), and 0.025 µg pTriEx™-2(Rluc). Mixture two contained 3 µl RiboJuice™ and 10 nM (final concentration) siRNA(b-gal)1. For each cell line, a reaction lacking siRNA was used as a control (set at 100%). Total volume per well was 300 µl. Cells were lysed with Reportasol™ Extraction Buffer 24 h after transfection and assayed for reporter activity. All assays were performed in triplicate and variation is expressed as standard error of the mean.

For more information on RiboJuice see inNovations 14, Article 3.

ProteoJuice™ Protein Transfection Reagent efficiently delivers protein into mammalian cells. The most rapid method for studying the effects of a specific protein in a mammalian cell is to transfect the protein directly into the cell. Introduction of a protein into cells permits studies of protein-protein interactions, protein turnover, cell signaling pathways, apoptotic pathways, and transcription factor-mediated gene regulation. ProteoJuice™ Protein Transfection Reagent is an effective reagent for the transfection of intact functional protein into mammalian cells.

Transfected histone localization in PC12 cells Cells were transfected with 1.25 µg Cy™ 3-conjugated histone H2A by using ProteoJuice. Images depict fluorescent (panel A) and brightfield (panel B) views of the same cells.

Transfected estrogen receptor localization in HepG2 cells HepG2 cells were transfected with FITC-labeled estrogen receptor by using ProteoJuice. After incubation, washed cells were exposed to DAPI nuclear stain for 2 min, followed by four additional washes in PBS before microscopy. Panels A, B, and C: no estradiol treatment; panels D, E, and F: estradiol (Calbiochem, 17b-estradiol) treatment. Images depict brightfield (panels A and D), FITC fluorescence (panels B and E), and DAPI fluorescence (panels C and F) views, with the same cells in panels A, B and C and in panels D, E, and F. Arrows indicate estrogen receptor localization in nuclei.

For more information on ProteoJuice see inNovations 17, Article 6.