DOTAP methosulfate

DOTAP or N-(2, 3-Dioleoyloxy-1-propyl) trimethylammonium methyl sulfate is a cationic liposome-forming compound for transfection of DNA, RNA and other negatively charged molecules into eukaryotic cells.

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.

Suitable for transient and stable transfection of DNA into cultured mammalian cells. Use approximately 5-10 μg DOTAP per μg DNA and approximately 10-20 μg DOTAP per mL cell culture medium. Protocol optimization within these guidelines provides very efficient transfection. The following cells have been successfully transfected using the DOTAP method:

A549
B6
BHK
BNL CL.2
C2
C6
Cardiomyocytes
CHO
COS-1
COS-7
Embryonic Fibroblasts (rat)
Embryonic stem cells (mouse)
Fibroblasts (primary human)
H225
Hepatocytes (primary rat)
HepG2
HEK293
HeLa
Hippocampal neurons (mouse and rat)
HMEC-1
Huh7
Jurkat
Lymphocytes (human peripheral blood)
M17
Macrophage (mouse)
MCF-7
MDA-MB-435
NIH3T3
Sf9
Smooth Muscle (rat aortic)
Spleen (mouse)
Vero
WI-38

A stable complex is formed when DOTAP is mixed with DNA in the absence of serum. The complexes are stable and can be directly added to the cell culture medium, where they fuse with the cell membrane, releasing the DNA into the cytoplasm. Note: complex formation is inhibited by serum, but once stable complexes have formed, the presence of serum is without consequence.