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.
|Published (Last):||4 August 2007|
|PDF File Size:||6.35 Mb|
|ePub File Size:||16.93 Mb|
|Price:||Free* [*Free Regsitration Required]|
An electroporation protocol for efficient DNA transfection in PC12 cells.
Identification by morphologic and immunologic criteria. This project was funded by the Cancer Society of New Zealand. 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. Toward development of artificial viruses for gene therapy: Articles from Journal of Biomolecular Techniques: However, it was observed in this study that transfection efficiencies appear to vary with some protocpl of HUVEC, and cells extracted from some cords are able to be transfected more efficiently than those from others.
Abstract Primary cells, such as HUVEC, are notoriously difficult to transfect and prltocol susceptible to the toxic effects of transfection reagents.
World J Gastroenterol ; Lipfoectamine were incubated for 5 min and then combined together for a further 20 min. Culture of human endothelial cells derived from umbilical veins.
Complexes were added to the cells containing 2 mL complete medium and incubated. Each point represents a different ratio of reagent: Gene Ther ; 5: Welsh S, Kay SA.
Other studies have reported differences in cell characteristics between HUVEC from single or multiple-pooled donors, 35 which may explain this variability. Physical methods of nucleic acid transfer: Infect Immun ; 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.
EGFP gene expression 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.
Reporter gene expression for monitoring gene transfer. Kreppel F, Kochanek S.
siRNA transfection in endothelial cells – siRNA, microRNA and RNAi
Differential ability of human endothelial cells to internalize and express exogenous DNA. Open in a separate window. Primary cells are considered more difficult to transfect than immortalized cell lines, as they are more susceptible to toxic agents and may degrade exogenous nucleic acids in the cytoplasm. Therefore, measurement of EGFP expression by flow cytometry may underestimate the total number of cells that was gene-modified initially.
Methods Mol Biol ; Transfection efficiency of cationic lipids with different hydrophobic domains in gene delivery. Nuclear envelope breakdown in mammalian cells involves stepwise lamina disassembly and microtubule-drive deformation of the nuclear membrane. Mol Ther ; 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 lipofectamihe use in subsequent assays.
Current developments in gene transfection agents. B Lipofectamine 2: Highly efficient transduction of endothelial cells by targeted artificial virus-like particles. Lipofectamine LTX was added, and the complexes were allowed to form by incubation for 25 min.