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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World J Gastroenterol ; Mixtures were incubated for 5 min and then combined together for a further 20 min. Representative traces of HUVEC, incubated for 48 h after transfection before analysis by flow cytometry.

Cationic liposomes induce macrophage apoptosis through mitochondrial pathway. Antibiotics and antifungal agents were not used during transfection procedures. Viral transduction is particularly efficient for gene transfer and is favored for in vivo use. Endothelial cell COX-2 expression and activity in hypoxia. Modification of adenovirus gene transfer vectors with synthetic polymers: Azzam T, Domb AJ.

Where a ratio was tested more than once, the mean transfection efficiency is plotted. Optimization of transfection of human endothelial cells. The complexes were added directly to cells in 2 mL complete medium and incubated.

An electroporation protocol for efficient DNA transfection in PC12 cells.

Curr Drug Deliv ; 1: Front Biosci ; 7: Efficient gene protoco into human umbilical vein endothelial cells allows functional analysis of the human tissue factor gene promoter. DNA was added to the mixture and incubated for a further 10 min. Methods Mol Protoco ; Medium was removed from the cells, replaced with complexes, and incubated.

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Abstract Primary cells, such as HUVEC, are notoriously difficult to transfect and are susceptible to the toxic effects of transfection reagents. RGS5, a hypoxia-inducible apoptotic stimulator in endothelial cells.

D Lipofectamine LTX 6. This study analyzed nine currently available, commercial transfection reagents and showed that lipofevtamine lipid reagents were the most efficient in gene-modifying HUVEC. Robinson1 and Gabi U.

The complexes were incubated on the cells for 3 h before 1 mL complete medium was added. Burlington, Ontario, Canada 9.

The complexes were added to cells in 2 mL complete medium and incubated. Angiogenic profiling and comparison of immortalized endothelial cells for functional genomics. Articles from Journal of Biomolecular Techniques: Transfection complexes were formed at room temperature in serum-free medium prior to drop-wise addition to HUVEC, followed by incubation for various periods and replacement with complete medium for 24 or 48 h.

Mean transfection efficiency for each reagent is shown horizontal bars. Identification by morphologic and immunologic criteria. Cell medium was replaced with 2 mL Opti-MEM I, to which the mixture was added, and incubated for 4 h, after which the complexes were replaced with complete medium.

An electroporation protocol for efficient DNA transfection in PC12 cells.

Therefore, measurement of EGFP expression by flow cytometry may underestimate the total proocol of cells that was gene-modified initially. Identification of replicative senescence-associated genes in human umbilical vein endothelial cells by an annealing control primer system. Exp Gerontol ; This led to an underestimation of the total amount of cell death caused by transfection but a more accurate representation of the number of live cells and proportion of EGFP-expressing cells left to plate out for subsequent assays.

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Cytokines Lipofwctamine Ther ; 2: Mol Ther ; 5: Green fluorescent protein as a novel tool to measure apoptosis and necrosis. Differences in EGFP expression were dependent mainly on transfection reagent.

siRNA transfection in endothelial cells – siRNA, microRNA and RNAi

A number of reasons have been considered to explain these differences. Dual targeting of gene delivery by genetic modification of adenovirus serotype 5 fibers and cell-selective transcriptional control.

Biotechnol Prog ; lipofectsmine Cancer Gene Ther ; 8: Welsh S, Kay SA. 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. There are no financial support or associations that would pose a conflict of interest. Each point represents a different ratio of reagent: Polymers for DNA delivery.