Product Name: Lipofectine

Cat No. : LF0001

Size: 1.5 ml

Product Description:

Lipofectine is a proprietary cationic lipid-based transfection reagent for mammalian cells. Change of cell culture media after DNA transfection is recommended but not required. 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 with minimal cytotoxicity. Change of cell culture media after DNA transfection is recommended but not required.

Protocol:

A. 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.

  B. 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.