70967
Reactivo de transfección GeneJuice®
Non-lipid based chemical transfection reagent optimized for maximum transfection efficiency, ease-of-use, and minimal cytotoxicity on a wide variety of mammalian cells.
Sinónimos:
Reactivo de transfección, Transfección GeneJuice
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About This Item
Nombre del producto
Reactivo de transfección GeneJuice®, Non-lipid based chemical transfection reagent optimized for maximum transfection efficiency, ease-of-use, and minimal cytotoxicity on a wide variety of mammalian cells.
form
liquid
manufacturer/tradename
Novagen®
storage condition
OK to freeze
technique(s)
transfection: suitable
shipped in
wet ice
storage temp.
2-8°C
Quality Level
Categorías relacionadas
Disclaimer
Features and Benefits
- Transferencia de ADN muy eficiente para transfecciones estables y transitorias
- Compatibilidad con medios que contienen suero y medios que no contienen suero
- Protocolo sencillo, sin necesidad de cambiar medios
- Ideal para transfección de gran rendimiento en formato de placa multipocillo
- Ideal para la producción de retrovirus para la transducción de células T
- Funciona para muchos tipos de células con una optimización mínima
- Proporciona una eficiencia de transfección mayor y una citotoxicidad menor que los reactivos de otros proveedores
- El tamaño de 1 ml proporciona suficiente reactivo como para realizar hasta 500 transfecciones en placas convencionales de 35 mm
General description
Stuart Rulten. Research Fellow. Universidad de Sussex
«Desde que trabajamos con GeneJuice, hemos ahorrado mucho tiempo y dinero»
Dr. Andrea Kress, Instituto de biología clínica y molecular, Erlangen, Alemania
«Me parece que Genejuice funciona para muchos tipos de células con una optimización mínima»
Caitriona Marie Lyons. Estudiante de doctorado University College Cork, Irlanda
La transfección es el proceso mediante el cual se introducen ácidos nucleicos en células de mamífero. El reactivo de transfección GeneJuice es una formulación patentada optimizada para una máxima eficiencia de la transfección, facilidad de uso y mínima citotoxicidad para las células de mamífero. Mientras que muchos de los reactivos de transfección disponibles se basan en la formulación lipídica catiónica, el reactivo de transfección GeneJuice está compuesto por una proteína celular no tóxica y una pequeña cantidad de una nueva poliamina. El reactivo de transfección GeneJuice permite una transferencia muy eficiente de ADN en transfecciones estables y transitorias de células eucariotas y es ideal para transfecciones de gran rendimiento en un formato de placa multipocillo. Su composición exclusiva es compatible con medios que contienen suero y medios que no lo contienen, lo que hace que sea innecesario el cambio de medios durante los experimentos de transfección. Genejuice es una alternativa superior a una amplia variedad de otras técnicas, como la coprecipitación de fosfato de calcio, la electroporación, la microinyección, la administración de partículas biolísticas, la lipofección y la formación de complejos con DEAE-dextrano. El tamaño de 1 ml proporciona suficiente reactivo para realizar hasta 500 transfecciones en placas convencionales de 35 mm.
Líneas celulares transfectadas con GeneJuice® Reactivo de transfección
| Líneas celulares: | células primarias: | ||||
|---|---|---|---|---|---|
| 10T1/2 293T 3T3 NIH 3T3 Swiss 3T3-L1 A204 A431 A549 alfa 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 Melanocito 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 | Células de músculo liso aórtico Astrocitos Angioblastos Condrocitos Células cromafines Células epiteliales: mamarias prostáticas traqueales Fibroblastos Queratinocitos |
Legal Information
Other Notes
Preparation Note
signalword
Danger
hcodes
Hazard Classifications
Eye Irrit. 2 - Flam. Liq. 2
Clase de almacenamiento
3 - Flammable liquids
wgk
WGK 1
flash_point_f
67.1 °F - Information taken from reference works and the literature.
flash_point_c
19.5 °C - Information taken from reference works and the literature.
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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.
Contenido relacionado
Successful delivery of nucleic acids and protein in eukaryotic cells requires a transfection protocol that maximizes transfection efciency, minimizes cytotoxicity, and results in the desired level of gene expression or silencing. EMD Millipore recognizes that there is not a one-size-ts-all solution to your transfection needs. We provide the breadth of transfection reagents necessary to give you the freedom to design the perfect experiment.
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