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.

Author: Dibar Kizahn
Country: Malawi
Language: English (Spanish)
Genre: Software
Published (Last): 16 August 2016
Pages: 414
PDF File Size: 20.43 Mb
ePub File Size: 8.3 Mb
ISBN: 218-7-29781-518-3
Downloads: 73579
Price: Free* [*Free Regsitration Required]
Uploader: Zulkizahn

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.

Highly efficient transduction of endothelial cells by targeted artificial virus-like particles. There are no financial support or associations that would pose a conflict of interest.

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. Comparison of the efficiency and safety of non-viral vector-mediated gene transfer into a wide range of human cells.

Complexes were incubated for 10 min.

Endothelial cell transfection with cationic liposomes and herpes simplex-thymidine kinase mediated killing. J Cell Sci ; Cell medium prohocol 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. This article has been cited by other articles in PMC.


Culture of human endothelial cells derived from umbilical veins. Angiogenic lpiofectamine and comparison of immortalized endothelial cells for functional genomics. Transfection efficiency of cationic lipids with different hydrophobic domains in gene delivery.

Front Biosci ; 7: Efficient gene transfer into human umbilical vein endothelial cells allows functional analysis of the human tissue factor gene promoter.

EGFP gene lipfoectamine allows easy determination of the proportion of cells that is gene-modified on a single-cell basis, detecting the number of cells expressing EGFP and their level of EGFP expression via flow cytometry.

Curr Opin Biotechnol ; 8: RGS5, a hypoxia-inducible apoptotic stimulator in endothelial cells.

siRNA transfection in endothelial cells – siRNA, microRNA and RNAi

An equal volume of PLUS reagent was added. Differences in EGFP expression were dependent mainly on transfection reagent. Identification by morphologic and immunologic criteria. Dachs, unpublished were used as negative and positive controls, respectively. The complexes were added to cells in 2 mL complete medium and incubated. Methods Mol Biol ; Towards endothelial-cell-directed pprotocol immunotherapy: Biochim Biophys Acta ; Therefore, measurement of EGFP expression by flow cytometry may underestimate the total number of cells that was gene-modified initially.

High-efficiency transient transfection of endothelial cells for functional analysis. A number of reasons have been considered to explain these differences.

An electroporation protocol for efficient DNA transfection in PC12 cells.

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 yield of transfected HUVEC for use in subsequent assays. Infect Immun ; Lipofectamine LTX was added, and the complexes were allowed to form by incubation for 25 min.


Polymers for DNA delivery. Curr Drug Saf ; 3: Inhibition of hydrophobic protein-mediated Candida albicans attachment to endothelial cells during physiologic shear flow.

Progress in developing cationic vectors for non-viral systemic gene therapy against cancer. 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. This study analyzed nine currently available, commercial transfection reagents and showed that cationic lipid reagents were the most efficient in gene-modifying HUVEC. Regulatory considerations for novel gene therapy products: DNA was added to the mixture and incubated for a further 10 min.

An electroporation protocol for efficient DNA transfection in PC12 cells.

Cytokines Mol Ther ; 2: Open in a separate window. Representative traces of HUVEC, incubated for 48 h after transfection before analysis by flow cytometry. This project lipofecgamine funded by the Cancer Society of New Zealand. Modification of adenovirus gene transfer vectors with synthetic polymers: 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 lttx Lipofectamine LTX showed no autofluorescence.

Comparison of nine transfection reagents. Exp Gerontol ; C Lipofectamine LTX 6. Currie1 Bridget A.