DOTAP chloride

A cationic lipid

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50mg

￥600.00

480.00

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100mg

￥1100.00

880.00

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250mg

￥2400.00

1920.00

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500mg

￥4037.00

3230.00

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货号: ajci67864
CAS: 132172-61-3
别名: (2,3-二油氧基丙基)三甲基氯化铵,1,2-Dioleoyl-3-trimethylammoniumpropane (chloride)
分子式: C42H80NO4?Cl
分子量: 698.6
纯度: >98%
溶解度: ≤33mg/ml in ethanol;0.5mg/ml in DMSO;5mg/ml in dimethyl formamide
储存: Store at -20°C, protect from light, stored under nitrogen
库存: 现货

Background

DOTAP chloride is a liposome formulations have been used to transfect plasmid DNA into eukaryotic cells in culture ^[1]. The synthetic cationic amphiphile DOTAP chloride has been shown to mediate uptake of plasmid also in the presence of a culture medium containing serum. It is metabolized in the cells and has proven to be less cytotoxic than other amphiphiles ^[2]. DOTAP chloride has also been used successfully for internalization of an 18-mer DNA oligonucleotide, and it was shown in these experiments to protect the oligonucleotide from degradation by nucleases in serum and cells ^[3].

参考文献:
[1]. Haukenes G, Szilvay A M, Brokstad K A, et al. Labeling of RNA transcripts of eukaryotic cells in culture with BrUTP using a liposome transfection reagent (DOTAP?)[J]. Biotechniques, 1997, 22(2): 308-312.
[2]. Leventis R, Silvius J R. Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles[J]. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1990, 1023(1): 124-132.
[3]. Capaccioli S, Dipasquale G, Mini E, et al. Cationic lipids improve antisense oligonucleotide uptake and prevent degradation in cultured cells and in human serum[J]. Biochemical and biophysical research communications, 1993, 197(2): 818-825.

DOTAP chloride 是一种脂质体制剂，已用于将质粒 DNA 转染到培养的真核细胞中 ^[1]。合成的阳离子两亲物 DOTAP chloride 已被证明在含有血清的培养基存在下也能介导质粒的摄取。它在细胞内代谢，已证明其细胞毒性低于其他两亲物 ^[2]。 DOTAP 氯化物也已成功用于 18 聚体 DNA 寡核苷酸的内化，并且在这些实验中表明它可以保护寡核苷酸免于被血清和细胞中的核酸酶降解^[3]。

Protocol

Cell experiment [1]:
Cell lines	mammalian CV-1 and 3T3 cells
Preparation Method	Lipid dispersions containing cationic lipid analogues were prepared by first bath-sonicating dried lipid samples in distilled water to clarity (2-5 min), then adding an equal concentration of 308 mM NaCl, 40 mM Hepes (pH 7.4), and sonicating further for 2 min. Lipid/DNA mixtures for transfections were prepared by incubating dotap chloride with DNA for 5 min, typically at a total lipid concentration of 0.5-2 mg/ml, before addition to cell monolayers. Reverse-phase evaporation vesicles were prepared as described previously [2] and filtered through 0.1/xm pore-size Nucleopore filters. Lipid mixing between cationic lipid dispersions and large unilamellar PC/PS vesicles was assayed by a resonance energy transfer-based procedure as described previously [3], incubating cationic lipid dispersions (2 μM), colabeled with 1 mol% (12-CPS)-18 PC and 0.35 mol% (12-DABS)-18 PC, with a 9-fold excess (18 μM) of unlabeled PC/PS vesicles. For transfection of cells using cationic lipid dispersions, cell monolayers were washed twice with HEPESbuffered saline, then incubated with lipid/DNA mixtures in the latter medium (total volume 3 ml per100-mm culture dish) at 37 °C, for 90 min except whereotherwise indicated. Following this incubation period,the incubation medium was either replaced or supplemented with 12 ml of D-MEM containing 5% serum,and the cells were cultured for a further 36 h beforeharvesting. Chloramphenicol acetyltransferase activityin plasmid pSV2cat-transfected cells was assayed asdescribed previously [4], using chloramphenicol andacetyl-CoA concentrations of 3.5 μM and 2.5 mM,respectively.
Reaction Conditions	0.5-2 mg/ml for 90 min
Applications	For transfection of the cells with a fixed amount of plasmid (usually 2 or 5 μg)
参考文献: [1]: Leventis R, Silvius J R. Interactions of mammalian cells with lipid dispersions containing novel metabolizable cationic amphiphiles[J]. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1990, 1023(1): 124-132. [2]: Wilschut J, Duzgunes N, Fraley R, et al. Studies on the mechanism of membrane fusion: kinetics of calcium ion induced fusion of phosphatidylserine vesicles followed by a new assay for mixing of aqueous vesicle contents[J]. Biochemistry, 1980, 19(26): 6011-6021. [3]: Silvius J R, Leventis R, Brown P M, et al. Novel fluorescent phospholipids for assays of lipid mixing between membranes[J]. Biochemistry, 1987, 26(14): 4279-4287. [4]: Gorman C M, Moffat L F, Howard B H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells[J]. Molecular and cellular biology, 1982, 2(9): 1044-1051.