Successful expression of recombinant proteins in mammalian or insect cells requires a transfection protocol that maximizes transfection efficiency, minimizes cytotoxicity, and results in the desired level of protein expression. EMD Millipore recognizes that there is not a one-size-fits-all solution to your transfection needs. We provide the breadth of transfection reagents necessary to give you the freedom to design the perfect experiment. Simplified protocols, including no media changes for most reagents and cell types, have been pre-optimized, requiring minimal assay development and allowing more time for research.
Successful Cell Transfection Strategies
Register now to participate in this complimentary webinar where you’ll learn how you can greatly improve your efficiency and increase protein expression in a few easy steps.
Adrian Vilalta, Ph.D.
Senior Manager, Molecular Biology R&D
EMD Millipore Corporation
November 6, 2012
NovaCHOice™ Transfection Kit
Chinese hamster ovary (CHO) cells are an excellent mammalian host system for transfection, recombinant protein expression and purification of authentically glycosylated and post-translationally folded proteins that resemble their natural counterparts. EMD Millipore’s NovaCHOice Kit provides optimal transfection in this widely utilized cell line. Quality controlled against suspension-adapted CHO cells, the NovaCHOice Kit is formulated to offer highly efficient DNA delivery with minimal cell death to maximize recombinant protein expression.
GeneJuice Performance ComparisonGeneJuice® transfection reagent provides higher protein expression levels than reagents from other suppliers.
Hela - SEAP Assay
Protein expression measured by Secreted Alkaline Phosphatase (SEAP) assay of HeLa cells at 24, 48 and 72-hour post transfection using GeneJuice® Transfection Reagent and five other commercially available reagents. At every time point, GeneJuice® Transfection Reagent shows a higher expression level than all the other products.
Result: GeneJuice® transfection reagent offers higher protein expression compared to reagents from other suppliers at all time points (24, 48 & 72 hrs post transfection in HeLa and CHO-K1 cells)
CHO-K1 - SEAP Assay
Protein expression measured by Secreted Alkaline Phosphatase (SEAP) assay of CHO-K1 cells at 24, 48 and 72-hour post transfection using GeneJuice® Transfection Reagent and five other commercially available reagents. At every time point, GeneJuice® Transfection Reagent shows a higher expression level than all the other products.
GeneJuice® transfection reagent provides higher transfection efficiency and lower cytotoxicity than reagents from other suppliers.
HeLa - 48-hr Post Transfection
CHO-K1-48-hr Post Transfection
|1.||Optimization using secreted alkaline phosphatase (SEAP) assay readout was performed to determine the optimal transfection conditions for each cell type (HeLa and CHO-K1) using GeneJuice® transfection reagent.|
|2.||Cells were plated in 500 µL medium and incubated at 37 ºC until they reached 60-70% confluency.|
|3.||Cells were transfected with a plasmid encoding GFP using various reagents according to their respective recommended protocols.|
|4.||Fluorescent photomicrographs (to visualize GFP expression) and phase contrast photomicrographs (to visualize morphology and health) were taken 48 hours post-transfection.|
- Overall, cells transfected with GeneJuice® transfection reagent, Reagent F6 & Reagent X9 showed normal morphology (even monolayer) 48-hr post transfection.
- While cells transfected with GeneJuice® reagent, Reagent F6 & Reagent X9 were healthier, GeneJuice® reagent provided the highest transfection efficiency among the three reagents.
- Cells transfected with Reagent L2K, Reagent JPE & Reagent JP were “rounded up”, indicating that they were under stress and were undergoing apoptosis.
- GFP expression was visible in those cells transfected using Reagent L2K, Reagent JPE & Reagent JP. But, as shown in the phase contrast photomicrographs, these cells did not exhibit normal morphology, indicating that they were under stress.
NanoJuice® Transfection KitEfficiently transfect difficult-to-transfect mammalian cell types with NanoJuice Transfection Kit, which is comprised of two separate reagents developed to work synergistically. Using the cutting-edge nanotechnology of Priostar® dendrimers along with a polycationic liposomal formulation, the NanoJuice reagent mixture provides excellent transfection rates, low cytotoxicity, and the methodological flexibility needed for primary cells and other difficult-to-transfect cells.
NanoJuice Transfection Reagent Is Less Cytotoxic Than Competitor Reagents
The indicated cell lines were plated in 24-well plates 18-24 h prior to transfection, such that cells were 80% confluent at time of transfection. Transfections were performed according to the manufacturers’ optimized protocols. For transfection, 0.25 µg of low endotoxin purified pTriEx-6 Rluc plasmid DNA was complexed with the relevant reagent and introduced into each well. After 24-48 h, the cells were extracted with Reportasol Extraction Buffer and Rluc activity was assayed. Data are represented as relative light units per well of 24-well plate (RLU/well). All values reflect an average of three replicate cultures.
NanoJuice Is Less Cytotoxic Than Competitor Reagents
Three replicate Saos-2 cultures were either left untreated, transfected with commonly used competitor's reagents, or transfected with NanoJuice Transfection Kit according to recommended protocols. Photographs were taken 48 h post-transfection.
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