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11202375001

Roche

DOTAP Liposomal Transfection Reagent

>99% (TLC), liquid, suitable for transfection

Synonym(s):

DOTAP methosulfate, liposomal transfection reagent, dotap, transfection reagent, dotap, N-(2,3-Dioleoyloxy-1-propyl)trimethylammonium methyl sulfate, Transfection reagent

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About This Item

Empirical Formula (Hill Notation):
C43H83NO8S
CAS Number:
Molecular Weight:
774.19
MDL number:
UNSPSC Code:
41106502
PubChem Substance ID:

description

N-[1-(2,3-Dioleoyloxy)propyl]-N,N,N-trimethylammonium methyl-sulfate

Quality Level

assay

>99% (TLC)

form

liquid

packaging

pkg of 5 × 400 μL (10 U/μl)

manufacturer/tradename

Roche

technique(s)

transfection: suitable

storage temp.

2-8°C

SMILES string

[H]C(C[N+](C)(C)C)(OC(CCCCCCC/C=C\CCCCCCCC)=O)COC(CCCCCCC/C=C\CCCCCCCC)=O.O=S([O-])(OC)=O

InChI

1S/C42H80NO4.CH4O4S/c1-6-8-10-12-14-16-18-20-22-24-26-28-30-32-34-36-41(44)46-39-40(38-43(3,4)5)47-42(45)37-35-33-31-29-27-25-23-21-19-17-15-13-11-9-7-2;1-5-6(2,3)4/h20-23,40H,6-19,24-39H2,1-5H3;1H3,(H,2,3,4)/q+1;/p-1/b22-20-,23-21-;

InChI key

RSMRWWHFJMENJH-LQDDAWAPSA-M

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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

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.
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.

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.

Storage Class

12 - Non Combustible Liquids

wgk_germany

nwg

flash_point_f

does not flash

flash_point_c

does not flash


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P Durante et al.
FEBS letters, 448(2-3), 239-243 (1999-04-28)
Fructose 2,6-bisphosphate is a potent endogenous stimulator of glycolysis. A high aerobic glycolytic rate often correlates with increased cell proliferation. To investigate this relationship, we have produced clonal cell lines of Rat-1 fibroblasts that stably express transgenes coding for 6-phosphofructo-2-kinase
In vivo OVA-specific Cytotoxic CD8+ T Cell Killing Assay
Nada C
Bio-protocol, 6 (2016)
B Weiss et al.
Cellular and molecular life sciences : CMLS, 55(3), 334-358 (1999-05-06)
Agents that produce their effects through an antisense mechanism offer the possibility of developing highly specific alternatives to traditional pharmacological antagonists, thereby providing a novel class of therapeutic agents, ones which act at the level of gene expression. Among the
I Knoke et al.
Human genetics, 104(3), 257-261 (1999-05-14)
Mutations in the androgen receptor gene (AR) cause a wide spectrum of androgen insensitivity syndromes (AIS). Mutation analysis of patients with AIS has revealed that the same missense mutation of the AR gene can give rise to strongly divergent phenotypes
V J Fincham et al.
Journal of cell science, 112 ( Pt 6), 947-956 (1999-02-26)
The v-Src oncoprotein perturbs the dynamic regulation of the cellular cytoskeletal and adhesion network by a mechanism that is poorly understood. Here, we have examined in detail the effects of a temperature-dependent v-Src protein on the regulation of p190 RhoGAP

Articles

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