Lipofectine
Catolog Number：LF0001
Features:
For DNA transfection to mammalian cells.
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.
Description:
Lipofectine is a proprietary cationic lipid-based transfection reagent for mammalian cells. The ratio of Lipofectine to plasmid DNA is fixed at 50 ul Lipofectine for 1 ug of plasmid DNA. Lipofectine 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 Lipofectine is consistently higher than that of similar product from other suppliers.
Protocol for transfection of adherent cells:
1. For each microgram of DNA to be transfected, add 50 ul of Lipofectine;
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 Lipofectine, change of media is not absolutely required.
Protocol for transfection of suspension cells:
Lipofectine has proven to be an effective reagent to transfect suspension cells. The ideal cell density at time of transfection is 1 x 106 per ml. Therefore, the day before transfection, suspension CHO cells are seeded at 0.5 x 106 per ml in media such as ProCHO (Lonza) or CD-CHO (Invitrogen) without serum. The amount of DNA used in transfection should be 3 μg 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.
