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GeneJuice® Transfection Reagent
Non-lipid based chemical transfection reagent optimized for maximum transfection efficiency, ease-of-use, and minimal cytotoxicity on a wide variety of mammalian cells.
别名:
GeneJuice Transfection, Transfection Reagent
About This Item
一般描述
Stuart Rulten. Research Fellow. University of Sussex
""Since we have been working with GeneJuice, we have saved a lot of time and money!""
Dr. Andrea Kress, Institute of Clinical and Molecular Virology, Erlangen, Germany
""I find that Genejuice works for a multitude of cell types with minimal optimization""
Caitriona Marie Lyons. PhD Student. University College Cork, Ireland
Transfection is the process by which nucleic acids are introduced into mammalian cells. GeneJuice Transfection Reagent is a proprietary formulation optimized for maximal transfection efficiency, ease of use, and minimal cytotoxicity for mammalian cells. Whereas many available transfection reagents are based on cationic lipid formulation, GeneJuice Transfection Reagent is composed of a nontoxic cellular protein and a small amount of a novel polyamine. GeneJuice Transfection Reagent enables highly efficient DNA transfer in both stable and transient transfections of eukaryotic cells and is ideal for high-throughput transfections in a multi-well plate format. The unique composition is compatible with both serum-containing and serum-free media, making media changes unnecessary during transfection experiments. Genejuice is a superior alternative to a wide variety of other techniques including calcium phosphate coprecipitation, electroporation, microinjection, biolistic particle delivery, lipofection, and complex formation with DEAE-dextran. The 1 ml size provides enough reagent to perform up to 500 transfections in standard 35 mm plates.
Cell lines transfected with GeneJuice® Transfection Reagent
| Cell Lines: | Primary Cells: | ||||
|---|---|---|---|---|---|
| 10T1/2 293T 3T3 NIH 3T3 Swiss 3T3-L1 A204 A431 A549 alpha TC1-6 AR 42J As4.1 AtT-20 B50 BC-1 BC-2 BC3 BCBL BHK-21 C3H/10T1/2 C6 C2C12 | Caco-2 Caki-1 Calpan-1 Calu-1 Calu-6 CCL-131 CFPAC-1 Chang Liver CHO CHO-7 CHO-IR CHO-K1 COS-1 COS-7 CS-1 CV-1 Daudi DDTI MF-2 DT40 ECV304 EL4 | ES-E14TG2a EVSCC17M H9c2 HCT-116 HEK293 HeLa HeLa B HeLa T4 Hep 3B2.1-7 HepG2 Hepa 1-6 Ht-29 HTB-37 HTB-45 Huh-7 HUVEC IC21 IEC-6 JEG-3 Jurkat KB | L57-3-11 L-6 L-929 MA-10 McA-RH7777 MCF-7 MCF-10-2A MDCK Melanocyte MG-63 Neuro 2A Neuroblastoma NRK NT2/D1 OV-1063 OVCAR3 P4 P19 PC12 PA317 PAM212 | PS-1 R2C RAW 264.7 RBL-2H3 RMP-41 SAOS-2 SC-1 Schneider line2 SK-N-MC SK-N-SH SKOV3 STO SW-480 SW-837 T3M4 TM4 U937 UCD Vero | Aortic smooth muscle cells Astrocytes Angioblasts Chondrocytes Chromaffin cells Epithelial cells: mammary prostate tracheal Fibroblasts Keratinocytes |
特点和优势
- Highly efficient DNA transfer for both stable and transient transfections
- Compatibility with both serum-containing and serum-free media
- Simple protocolno need for media changes
- Ideal for high-throughput transfection in a multi-well plate format
- Ideal for retrovirus production for T-cell transduction
- Works for a multitude of cell types with minimal optimization
- Provides higher transfection efficiency and lower cytotoxicity than reagents from other suppliers
- The 1 ml size provides enough reagent to perform up to 500 transfections in standard 35 mm plates
警告
制备说明
其他说明
法律信息
警示用语：
Danger
危险声明
危险分类
Eye Irrit. 2 - Flam. Liq. 2
WGK
WGK 1
闪点(°F)
Information taken from reference works and the literature.
闪点(°C)
Information taken from reference works and the literature.
Why do I need to perform optimization? How should I go about doing it?
Each cell type behaves differently, by carrying out an optimization, the best transfection condition for your particular cell type can be determined. In other words, you can avoid putting too much transfection reagent on your cells, which may cause unnecessary toxicity issue and waste of precious transfection reagent. Optimization is suggested for every new combination of cell type and plasmid. The most important parameters are cell density and ratio of transfection reagent to DNA. Start with the volume of the selected transfection reagent (1x) and plasmid amount (1x) as recommended in the User Protocol. If those conditions do not yield the desired results, an optimization experiment can be performed. In a 24-well plate, plate the same amount of cells in each well. Set up a gradient across the plate and add the appropriate volume of transfection reagent (0.5x, 1x, 1.5x, 2x, 2.5x and 3x). Set up a gradient down the plate and add the appropriate amount of plasmid (0.5x, 1x, 1.5x and 2x). With a reporter gene in the plasmid, the optimal condition can be easily determined.
What is the size limit for plasmid DNA?
Large plasmids in the range of 12-15 kb can be transfected. We have cloned and expressed inserts encoding large proteins (including β-gal) without difficulty in mammalian cell lines.
Is the quality of DNA important for good transfection?
Yes, it is essential that the DNA to be transfected is of high quality and free of endotoxins. Plasmid DNA preparations should include an endotoxin removal step.
How long should I leave the transfection reagent on the cells? Do I need to change medium at any time after transfection?
Since our nucleic acid transfection reagents are compatible with serum-containing media, medium change after transfection is not necessary. The majority of cell types can be incubated with the transfection mix for 24-72h without any media change, and then harvested for the desired downstream application. If media change is necessary due to the toxicity of the protein being expressed, the transfection mixture can be removed after 2-8 h of incubation and replaced with complete growth medium.
Can I use the product in the presence of serum?
Yes. Our nucleic acid transfection reagents are effective for transfecting cells in media with or without serum. While cells can be incubated in media containing serum, it is absolutely critical that serum is NOT present during formation of the transfection reagent/DNA complex. For most applications, we recommend adding the transfection reagent/DNA complex (formed in serum-free media) to cells grown in complete growth media. For certain cell lines and experimental conditions, serum starvation of cells might be required. Since serum provides growth factors and nutrients, transfection efficiencies achieved with growth in serum containing media are typically better than those in serum-free media.
Can the DNA transfection reagents be used for co-transfecting plasmids?
Yes. Multiple plasmids can be transfected into the cell at the same time. The key is to maintain the optimal ratio of total DNA (all plasmids). See the User Protocols for more information on the ratio of reagent to DNA.
Will antibiotics interfere with transfection?
We do not recommend including antibiotics during the formation of the transfection reagent/DNA complex. Increased cell permeability during transfection causes high antibiotic influx, resulting in cell death. Some antibiotics (such as kanamycin) are cationic and can therefore interfere with transfection. Antibiotics such as penicillin and streptomycin can be present in the complete growth media (with serum) which is used to grow the cells. If you are generating stable transfectants, add selection antibiotics (e.g., G 418 or hygromycin) 48-72h after transfection.
How do I scale my transfection protocol when working with different culture volumes?
For most standard culture formats, guidelines are provided in the User Protocol. If you are using different culture volumes, vary the amounts of DNA, transfection reagent, cells, and culture media in proportion to the relative surface area while keeping the transfection reagent: DNA ratio constant.
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