Transfection

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Merck:/Freestyle/BI-Bioscience/Genomic-Analysis/transfection-cover.jpgTransfection: A Science and an Art.
Optimized Reagents for Efficient Transfection.

Transfection Reagents for Expression of Recombinant Proteins

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. MilliporeSigma 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.

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What Customers Are Saying About GeneJuice® Reagent...

“I use GeneJuice® reagent because of its ease-of-use, low toxicity and wide range of target cells.”
Stuart Rulten, Research Fellow, University of Sussex

“Since we have been working with GeneJuice® reagent, we have saved a lot of time and money!”
Dr. Andrea Kress, Institute of Clinical and Molecular Virology, Erlangen, Germany

“I find that Genejuice® reagent works for a multitude of cell types with minimal optimization.”
Caitriona Marie Lyons, PhD Student, University College Cork, Ireland

Transfection Reagent
Cell Lines and Applications
GeneJuice® Reagent The standard for transfection in most cell lines in serum-containing or serum-free media
NovaCHOice® Kit Efficient, scalable protein expression in suspension CHO lines
293-Free™ Reagent Maximal output from HEK293 suspension cultures
NanoJuice® Kit Overcome the most difficult-to-transfect mammalian cells (e.g. primary cell lines)
Insect GeneJuice® Reagent Optimized for insect cells
RiboJuice™ Reagent Effective delivery of siRNA for targeted gene silencing in most cell lines
ProteoJuice™ Reagent Delivery of even the most complex proteins into most cells

GeneJuice® Reagent Performance Comparison

GeneJuice® reagent transfection reagent provides higher protein expression levels than reagents from other suppliers.

Hela - SEAP Assay

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.
CHO-K1 - SEAP Assay

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.

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-K1cells)


GeneJuice® Transfection Reagent Provides Higher Transfection Efficiency and Lower Cytotoxicity than Reagents from Other Suppliers.

HeLa - 48-hr Post Transfection

HeLa - 48-hr Post Transfection
CHO-K1-48-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. Result:
  • 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.


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. $’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.

Detect protein expression earlier when you transfect with the NovaCHOice® Transfection Kit
Detect protein expression earlier when you transfect with the NovaCHOice® Transfection Kit.
b-galatosidase expression data in CHO-S cells at 1, 3, 5 and 7 days post transfection using the NovaCHOice® Transfection Kit and a competing reagent according to manufacturer‘s protocol. Results show that higher protein expression was detected on Day 1 in cells transfected with NovaCHOice® Transfection Kit than in those transfected with the Transfection Reagent F.


NanoJuice® Transfection Kit

Efficiently 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
 
Efficiently transfect difficult-to-transfect mammalian cell types with NanoJuice® Transfection Kit
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

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.


Antibiotics and Selection Agents

To select cells that have been successfully transfected with DNA carrying an antibiotic resistance marker, choose from our wide variety of high quality Calbiochem cell-culture tested antibiotics. Cited in thousands of peer-reviewed journals, Calbiochem antibiotics are manufactured in a controlled environment and perform with exceptional consistency to ensure the best results, lot after lot. Examples of Calbiochem antibiotics frequently cited in publications:

Antibiotic
Catalog Number
# of Citations from 2000-2009
G-418 Sulfate, cell culture tested 345810 6200
Bafilomycin A1, Streptomyces griseus 196000 460
Hygromycin B, Streptomycessp., cell culture tested 400052 2550