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D1163 Sigma

DOTAP methosulfate

Reagent for stable and transient DNA/RNA transfections

Synonym: N-(2,3-Dioleoyloxy-1-propyl)trimethylammonium methyl sulfate, Transfection reagent

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Properties

Related Categories Cell Biology, Cell Signaling and Neuroscience, Cloning and Expression, Molecular Biology, Signal Transduction Research Tools,
grade   for molecular biology
InChI Key   RSMRWWHFJMENJH-LQDDAWAPSA-M
form   liquid (MES buffered saline)
usage   sufficient for 40 transfections (10 cm dishes)
concentration   1 mg/mL
color   clear to hazy colorless to faint yellow
storage temp.   2-8°C

Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

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.

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

Features and Benefits

• Up to 100x more efficient than calcium phosphate and DEAE transfection
• Lower toxicity than calcium phosphate and DEAE based transfection
• Compatible with both serum and serum-free transfection protocols
• Compatible with in-vivo transfection protocols

Principle

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.

General description

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.

Price and Availability

Safety & Documentation

Safety Information

Personal Protective Equipment 
RIDADR 
NONH for all modes of transport
WGK Germany 
3

Documents

Certificate of Analysis

Certificate of Origin

Protocols & Articles

Articles

Introduction to Transfection

Transfection involves the introduction of DNA or RNA into eukaryotic cells and is used in research to study and modulate gene expression. Thus, transfection techniques serve as an analytical tool tha...
Keywords: Apoptosis, Biological techniques, Drug discovery, Gene expression, Genetic, Precipitation, Transfection, transformation

Transfection Best Practices

The work of scientific research is an art which requires good attention to detail.  The quality of transfection is highest when these commonly used best practices and notes on optimization are employ...
Keywords: Antibiotics, Apoptosis, Growth factors, Molecular biology, Purification, Transfection

Transfection Reagent Selection Guide

Choosing a transfection reagent is easier when you can rely on the work of researchers who have used it before you. During development of each reagent, good transfection efficiency was verified in a ...
Keywords: Transfection

Protocols

Calcium Phosphate Transfection Kit Protocol

Calcium phosphate transfection is a commonly used method for the introduction of DNA into eukaryotic cells. This technique has been used to obtain both transient1 and stable2 transfections in a wide ...
Keywords: Antibiotics, Applications, Cell culture, Gene expression, Molecular biology, Transfection, Type

Escort™ III Transfection Reagent Protocol

Escort III is a liposome suspension composed of a polycationic lipid and a neutral, non-transfecting lipid compound.  Transfection using liposomes is a commonly used method for the introduction of DN...
Keywords: Cell culture, Transfection

Escort™ IV Transfection Reagent Protocol

Escort IV is a liposome suspension composed of a polycationic lipid and a neutral, non-transfecting lipid compound.  Transfection using liposomes is a commonly used method for the introduction of DNA...
Keywords: Cell culture, Transfection

Lipid Transfection Optimization Protocol

There are many steps of the lipid transfection protocol that can be optimized if the initial transfection attempt is not successful. You may boost transfection efficiency and/or lower toxicity by opt...
Keywords: Cell culture, Transfection

Peer-Reviewed Papers
15

References

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