lipo 2000说明书.docxVIP

Lipofectamine? 2000转染说明书IntroductionLipofectamine? 2000 CD is a proprietary animal origin-free formulation for the transfection of nucleic acids into eukaryotic cells. A Drug Master File (DMF) has been submitted to the FDA. Contact Technical Service or your local Sales Representative for permission to cross-reference the DMF. Using Lipofectamine? 2000 CD for transfection provides the following advantages: Highest transfection efficiency in many cell types and formats. Refer to the Cell Lines database at for a list of cell typestransfected. he animal origin-free formulation ensures that mammalian cell culture and bioproduction processes are free of animal-derived materials. DNA-Lipofectamine? 2000 CD complexes can be added directly to cells in culture medium. It is not necessary to remove complexes or change/add medium after transfection, but complexes may be removed after 4-6 hours. Important Guidelines for TransfectionCulture cells in serum-free media that are free of animal-derived components. Test serum-free media for compatibility with Lipofectamine? 2000 CD since some serum-free formulations (e.g. CD 293, 293 SFM II, CD Hybridoma) may inhibit cationic lipid-mediated transfection. For consistent animal origin-free transfection, use OptiPro? SFM (Cat. No. 12309-019) to dilute DNA and Lipofectamine? 2000 CD before complexing. Transfect cells at high cell density: For adherent cells, 90-95% confluence at the time of transfection is recommended for high efficiency, high expression levels, and to minimize cytotoxicity. For suspension cultures, transfect cells at a density of 0.8-1.1 x 106 cells/ml. Optimization may be necessary. Maintain the same seeding conditions between experiments. Do not add antibiotics to media during transfection as this causes cell death. Do not add Pluronic? or charged media extracts (e.g. dextran sulfate) to media during transfection as these reagents can inhibit transfectionTransfecting Adherent CellsUse the following procedure to transfe