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DNA Transfection Kit
Each cell line and plasmid is different. The protocols and recommendations are for CHO cells. Investigators should optimize the condition for their cell lines and plasmids.
MB088-450-20: Safectine RU50 DNA Transfection Kit
Safectine RU50 DNA Transfection Kit is a proprietary cationic lipid-based transfection reagent for mammalian cells. The ratio of Safectine RU50 to plasmid DNA is fixed at 50 ul Safectine RU50 for 1 ug of plasmid DNA. Safectine RU50 DNA Transfection Kit is formulated in a ready-to-use format - all users need to do is add DNA and vortex the mix. After a 15-minute incubation, the transfection mix can be added to cells. As judged by target protein production, the transfection efficiency of Safectine RU50 is consistently higher than that of similar product from other suppliers.
1. Simplest transfection protocol - ready to use, just mix with DNA and vortex;
2. Superior transfection efficiency;
3. Effective for both adherent and suspension cells;
4. Minimal cytotoxicity;
5. Change of cell culture media after transfection is recommended but not required;
6. Highly reproducible results;
7. Product stable at 4°C and at -20°C.
Protocol for transfection of adherent cells:
1. For each microgram of DNA to be transfected, add 50 ul of Safectine RU50;
2. Flick, rock, pipet up and down, or vortex the mix for 5 seconds;
3. Incubate the mix at room temperature for 15 minutes;
4. Add the mix to media and cells.
Note: Change of media 12 to 24 hour post transfection may improve cell growth condition, and is recommended. With Safectine, change of media is not absolutely required.
Protocol for transfection of suspension cells:
Safectine has proven to be an effective reagent to transfect suspension cells. The ideal cell density at time of transfection is 1 x 10^6 per ml. Therefore, the day before transfection, suspension CHO cells are seeded at 0.5 x 10^6 per ml in media such as ProCHO (Lonza) or CD-CHO (Invitrogen) without serum. The amount of DNA used in transfection should be 3 ug per ml of cells, which yields optimal cell growth and target protein expression. Transfection mix can be prepared exactly as described in the previous section. The transfection mix is then added to the suspension cells. Media replacement is recommended 12 to 24 hour post transfection, and data has shown that media replacement can increase cell growth rate after transfection for suspension. For protein or antibody production, cell media can be harvested 4-7 days post transfection.
Note: Each cell line and plasmid is different. The protocols and recommendations are for CHO cells. Users should optimize the condition for their cell lines and plasmids.
1. "CHO-K1 cells were transfected with Safectine RU50 (Syd Labs) according to the manufacturer's protocol.". Wu Z, Li X, Sunkara M, Spearman H, Morris AJ, Huang C. PIPKIgamma Regulates Focal Adhesion Dynamics and Colon Cancer Cell Invasion. PLoS One. 2011; 6(9):e24775.
2. "or transfection, the different expression plasmids were introduced into HEK293 cells using Safectine RU50 DNA transfection kit according to the manufacturer's protocol (Syd Labs, Malden, MA, USA).". Dong G, Gross K, Qiao F, Ferguson J, Callegari EA, Rezvani K, Zhang D, Gloeckner CJ, Ueffing M, Wang H. Calretinin interacts with huntingtin and reduces mutant huntingtin-caused cytotoxicity. J Neurochem. 2012 Nov;123(3):437-46.
3. "CHO-K1 and 293T cells were transfected with Safectine RU50 according to the manufacturer's protocol." Li X, Zhou Q, Sunkara M, Kutys ML, Wu Z, Rychahou P, Morris AJ, Zhu H, Evers BM, Huang C. Ubiquitylation of phosphatidylinositol 4-phosphate 5-kinase type I γ by HECTD1 regulates focal adhesion dynamics and cell migration. J Cell Sci. 2013 Jun 15;126(Pt 12):2617-28.
4. "For transfection, the different expression plasmids were introduced into HEK293 cells using a Safectine RU50 DNA transfection kit according to the manufacturer's protocol (Syd Labs)." Dong G, Gross K, Qiao F, Ferguson J, Callegari EA, Rezvani K, Zhang D, Gloeckner CJ, Ueffing M, Wang H. Calretinin interacts with huntingtin and reduces mutant huntingtin-caused cytotoxicity. J Neurochem. 2012 Nov;123(3):437-46.