Description
General description
DOTAP Liposomal Transfection Reagent is a liposome formulation of the cationic lipid DOTAP. Unlike other liposomal reagents, it produces highly efficient transfections both in the presence and absence of serum. It can also be successfully used for in vivo applications. Mixing DOTAP with DNA results in spontaneously formed stable complexes. These complexes can be added directly to the cell culture medium. This method of DNA transfer is very gentle, avoiding the cytotoxic effects commonly encountered with lipofection or other transfection methods.
Application
Cationic liposome-forming compound for transfection of DNA, RNA and other negatively charged molecules into eukaryotic cells. Mixing DOTAP with DNA results in spontaneously formed stable complexes that can be directly added to cell culture medium. These complexes fuse with the cell membrane and release DNA into the cytoplasm. The cells are transfected efficiently without cytotoxic effects. Use approximately 5-10 μg DOTAP per μg DNA and approximately 10-20 μg DOTAP per mL cell culture medium. The optimal working concentration and transfection time depends in part on the cell line used, and the type of material (RNA, DNA, protein) loaded.
The DOTAP (N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate) Liposomal Transfection Reagent is a liposome formulation of the cationic lipid DOTAP. DOTAP can be used for the highly efficient transfection of DNA including yeast artificial chromosomes (YACs) into eukaryotic cells for transient or stable gene expression, and is also suitable for the efficient transfer of other negatively charged molecules, such as RNA, oligonucleotides, nucleotides, ribonucleo-protein (RNP) complexes, and proteins into research samples of mammalian cells.
Features and Benefits
Easy to use: The lipid dispersion is simply mixed with the DNA solution, then applied directly to the cells
Highly effective: 5 to 100-fold more effective than the calcium phosphate or DEAE-dextran method
Gentle: No cytotoxic effects
Flexible: Equally effective in the presence or absence of serum. Effective on a wide range of species (e.g., insect cells, mammalian cells) and a variety of different cell types. Can be used for either transient or stable transfection procedures.
Aqueous dispersion (liposomes) in MBS (MES-buffered saline, pH 6.2) (bottled under argon), 1mg/ml, sterile-filtered.
Preparation Note
Working concentration: Approximately 5 - 10μg DOTAP/μg DNA, and 10 - 20μg DOTAP/ml cell culture medium.
The optimal working concentration is dependent on several parameters, including the cell line being used, concentration and type of nucleic acid (DNA, RNA), and incubation time. It is important to optimize the transfection conditions for the individual cell type studied.
Cytotoxicity: Not cytotoxic up to a concentration of 100μg/ml (PBLs, HeLa cells).
Other Notes
For life science research only. Not for use in diagnostic procedures.
