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 2-8h of incubation and replaced with complete growth medium.
72181
293-Free Transfektionsreagens
Animal-free polycationic liposomal transfection reagent optimized for the transfection of HEK293 cells grown in suspension culture.
Synonym(e):
Gene delivery
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| Größe/SKU | Verfügbarkeit | Preis |
|---|---|---|
1 mL | Warenkorb auf Verfügbarkeit prüfen | € 582,00 |
5 mL | Warenkorb auf Verfügbarkeit prüfen | € 2.410,00 |
10 mL | Warenkorb auf Verfügbarkeit prüfen | € 4.320,00 |
Über diesen Artikel
NACRES:
NA.54
UNSPSC Code:
41105500
eCl@ss:
32039090
Form:
liquid
Technique(s):
transfection: suitable
€ 582,00
Warenkorb auf Verfügbarkeit prüfen
Technischer Dienst
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Unterstützung erhaltenQuality Segment
form
liquid
manufacturer/tradename
Novagen®
storage condition
OK to freeze
technique(s)
transfection: suitable
shipped in
wet ice
storage temp.
2-8°C
General description
Humane embryonale 293-Nierenzellen, oft auch als HEK 293, HEK-293 oder 293-Zellen bezeichnet, sind spezifische Zelllinien, die ursprünglich aus humanen embryonalen Nierenzellen gewonnen und in Gewebekultur gezüchtet wurden. Aufgrund ihres robusten Wachstums werden HEK293-Zellen oft für Forschungsanwendungen im Bereich der Zellbiologie mit DNA-Plasmiden transfiziert. Außerdem werden HEK293-Zellen von Biotechnologieunternehmen zur Herstellung von therapeutischen Proteinen und Viren für die Gentherapie verwendet. Das Transfektionsreagenz 293-Free<TMSYMBOL></TMSYMBOL> besteht aus einer einzigartigen polykationischen liposomalen Formulierung und wurde speziell für die Transfektion von in Suspensionskultur gewachsenen HEK293-Zellen entwickelt. Es eignet sich hervorragend für die Proteinproduktion von Säugern. Das aus nicht-tierischen Quellen gewonnene Transfektionsreagenz 293-Free weist nur minimale Zelltoxizität auf. Es wird als sterile und gebrauchsfertige Lösung geliefert. Die 1-mL-Packungsgröße bietet ausreichend Reagenz für die Transfektion von einem Liter Kultur.
Polykationisches liposomales Transfektionsreagenz ohne tierische Bestandteile, optimiert für die Transfektion von in Suspensionskultur gewachsenen HEK293 Zellen.
Features and Benefits
- Hervorragende Transfektionseffizienz für HEK293-Suspensionskulturen zur Maximierung der Proteinexpressionswerte
- Aus nicht-tierischen Quellen gewonnen, um den Übergang vom Forschungsstadium in die Entwicklungsphase zu beschleunigen
- Vereinfachtes Protokoll ohne Medienwechsel, aufgrund der Kompatibilität sowohl mit serumhaltigen als auch serumfreien Medien
- Minimale Zelltoxizität führt zu höheren Proteinexpressionswerten
Other Notes
Aufgrund der Natur der in dieser Lieferung enthaltenen Gefahrgüter können zusätzliche Versandkosten erhoben werden. Bestimmte Größen sind ggf. von den Versandzuschlägen für Gefahrgüter ausgenommen. Um weitere Informationen über diese Versandzuschläge zu erhalten, kontaktieren Sie bitte Ihren lokalen Vertrieb.
Legal Information
NOVAGEN is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxizität: Entzündbar (J)
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Dieser Artikel | |||
|---|---|---|---|
| description Animal-free polycationic liposomal transfection reagent optimized for the transfection of HEK293 cells grown in suspension culture. | description Efficient protein delivery into mammalian cells | description Optimized transfection reagent specifically developed for mammalian protein production in chinese hamster ovary (CHO) cells. | description Novagen® |
| technique(s) transfection: suitable | technique(s) transfection: suitable | technique(s) transfection: suitable | technique(s) - |
| form liquid | form liquid | form - | form liquid |
| Quality Level 200 | Quality Level 100 | Quality Level 100 | Quality Level 100 |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. −20°C | storage temp. −20°C |
| shipped in wet ice | shipped in - | shipped in - | shipped in wet ice |
| storage condition OK to freeze | storage condition OK to freeze | storage condition OK to freeze | storage condition OK to freeze, avoid repeated freeze/thaw cycles |
Signalwort
Danger
Gefahrencodes
Sicherheitshinweise
Hazard Classifications
Eye Irrit. 2 - Flam. Liq. 2
Lagerklasse
3 - Flammable liquids
WGK
WGK 2
Flammpunkt (°F)
67.1 °F - Information taken from reference works and the literature.
Flammpunkt (°C)
19.5 °C - Information taken from reference works and the literature.
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How long should I leave the transfection reagent on the cells? Do I need to change medium at any time after transfection?
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Can the DNA transfection reagents be used for co-transfecting plasmids?
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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.
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Why do I need to perform optimization? How should I go about doing it?
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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.
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How important is the cell density/confluency and cell passage number at the time of transfection?
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Transfection of nucleic acids requires cells to be actively dividing. Therefore, the optimum cell density for transfection is generally between 50-80% confluency for adherent cells and 1.0-2.0 x 106 cells/mL for suspension cells. Avoid using extensively passaged cells; use a fresh vial of cells if a sudden drop in transfection efficiency is noticed. If necessary, determine the optimum cell density and passage number range for every new cell line, and keep these parameters constant for all experiments to ensure reproducibility.
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What is the size limit for plasmid DNA?
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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.
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Will antibiotics interfere with transfection?
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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.
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Is the quality of DNA important for good transfection?
1 answer-
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.
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How do I scale my transfection protocol when working with different culture volumes?
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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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Can I use the product in the presence of serum?
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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.
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