LIPOFECTAMINE LTX PROTOCOL PDF

Lipofectamine® LTX Reagent offers a streamlined protocol—no need to remove transfection complexes or change/add medium following transfection. A simple. Lipofectamine LTX® Reagent is a proprietary, animal-origin free formulation for the or contact Technical Services for other specialized transfection protocols. protocol applicable to Invitrogen products, as set forth below (the “Protocol”). by adding 50 μL of Lipofectamine™ LTX to μL of Opti-MEM® medium.

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Arch Biochem Biophys ; This project was funded by the Cancer Society of New Zealand. Currie1 Bridget A. Open in a separate window. Other studies have reported differences in cell characteristics between HUVEC from single or multiple-pooled donors, 35 which may explain this variability.

Lipofectamine LTX was identified as the optimal transfection reagent as a result of its higher transfection efficiency at shorter periods of time following transfection when cytotoxicity was limited, allowing sufficient lipofectamjne of transfected HUVEC for use in subsequent assays.

An electroporation protocol for efficient DNA transfection in PC12 cells.

Increasing the time to analysis from 24 h to 48 h resulted in an increased proportion of EGFP-positive cells for some reagents but also caused a reduction in cell viability. The complexes were added to cells in 2 mL complete medium and incubated. BoxChristchurchNew Zealand Phone: This article has been cited by other articles in PMC. The proportion of EGFP-positive cells is presented on the graphs. Abstract Primary cells, such as HUVEC, are notoriously difficult to transfect and are susceptible to the toxic effects of transfection reagents.

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An equal volume of PLUS reagent was added. Hum Gene Ther ; For three of the reagents Effectene, Escort IV, and ExGenthe final amount of DNA played a role in transfection efficiency, with more DNA in the mixture associated with increased transfection efficiency results not shown.

lipofectamihe Nuclear envelope breakdown in mammalian cells involves stepwise lamina disassembly and microtubule-drive deformation of the nuclear membrane. Cytokines Mol Ther ; 2: A transfection reagent with a high transfection efficiency and low cytotoxicity was sought to retain sufficient viability of transfected HUVEC for subsequent assays. High-efficiency transient transfection of endothelial cells for functional analysis.

DNA was added to the mixture and the complexes incubated for 15 min.

On the other hand, luciferase activity, detected via conversion of a substrate, resulting in amplified signal, determines the behavior of the entire population, thereby losing information about single cells.

Br J Haematol ; DNA was added to the mixture and incubated for a further 10 min.

An electroporation protocol for efficient DNA transfection in PC12 cells.

Transfer and expression of foreign genes in mammalian cells. B Lipofectamine 2: Regulatory considerations for novel gene therapy products: Dual targeting of gene delivery by genetic modification of adenovirus serotype 5 fibers and cell-selective transcriptional control.

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L—L [ PubMed ]. It has been reported that some cationic liposome transfection reagents could lead to autofluorescence in fluorescent microscopy and flow cytometry analysis, 38 but our results for mock transfection using Lipofectamine and Lipofectamine LTX showed no autofluorescence. Kreppel F, Kochanek S. Robinson1 and Gabi U. Differential ability of human endothelial cells to internalize and express exogenous DNA.

siRNA transfection in endothelial cells – siRNA, microRNA and RNAi

Reporter gene expression for monitoring gene transfer. Representative traces of HUVEC, incubated for 48 h after transfection before analysis by flow cytometry.

Optimization of transfection of human endothelial cells. Burlington, Ontario, Canada 9. There are no financial support or associations that would pose a conflict of interest. Efficient gene transfer into human umbilical vein endothelial cells allows functional analysis of the human tissue factor gene promoter.

Non-viral gene delivery with cationic liposome-DNA complexes. Cells were incubated for lipofectaimne h, after which, the complexes were replaced with complete medium.

Identification by morphologic and immunologic criteria. Cancer Gene Ther ; 8: