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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An electroporation protocol for efficient DNA transfection in PC12 cells.
We thank Sigma-Aldrich, Fermentas, and Roche, which kindly supplied free samples of their transfection reagents for us to test. Mol Ther ; 5: Antibiotics and antifungal agents were not used during transfection procedures. Improving safety of gene therapy.
Pdotocol Mol Biol ; The length of incubation prior to analysis made a difference in four of the nine reagents tested, and higher expression levels were detected after 48 h compared with 24 h Fig. DNA was added to the mixture and the complexes incubated for 15 min. 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.
This study analyzed nine currently available, commercial transfection reagents and showed that cationic lipid reagents were the most efficient in gene-modifying HUVEC.
Biochim Biophys Acta ; Curr Drug Deliv ; 1: A number of reasons have been considered to explain these differences. Chemical transfection reagents have been shown to reduce growth and viability of cells after transfection, possibly as a result of changes in the strength of the cell membrane.
On the other hand, luciferase activity, detected via conversion of a substrate, resulting in amplified signal, determines the behavior of the entire population, thereby ltd information about single cells.
Physical methods of nucleic acid transfer: Curr Drug Saf ; 3: Viral transduction is particularly efficient for gene transfer and is favored for in vivo use.
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Gene Ther ; 5: 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.
Lipfectamine Opin Biotechnol ; 8: Hum Gene Ther ; High-efficiency transient transfection of endothelial cells for functional analysis.
Cells can be gene-modified in vitro and in vivo using physical, viral, or chemical methods. Mol Ther ; Our study demonstrated that a small selection of commercially available chemical transfection reagents was able to transfer exogenous genes efficiently to primary human cells.
Villemejane J, Mir LM. DNA was added to the mixture and incubated for a further 10 min. J Biol Chem ; 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.
Flow cytometry black, AAD-positive cells indicated that cells analyzed after 48 h tended to have more dead cells compared with cells analyzed after 24 h, e.
siRNA transfection in endothelial cells – siRNA, microRNA and RNAi
Nonviral approaches for targeted delivery of plasmid DNA ;rotocol oligonucleotide. Other studies have reported differences in cell characteristics between HUVEC from single or multiple-pooled donors, 35 which may explain this variability. Cells were incubated for 3 h, after which, the complexes were replaced with complete medium. Transductional and transcriptional targeting of adenovirus for clinical applications.
Lipofectamkne targeting of gene delivery by genetic modification of adenovirus serotype 5 fibers and cell-selective transcriptional control.
Green fluorescent protein as a novel tool to measure apoptosis and necrosis. There are no financial support or associations that would pose a conflict of interest. The nine compounds tested in this study included activated dendrimers, cationic polymers of linear PEI, lipids and polyamines, nonliposomal lipids, polycationic lipids, and cationic lipids, reflecting the broad categories of chemical transfection reagents available and the intense development in this area.
The complexes were added directly to cells in 2 mL complete medium and incubated. Br J Pharmacol ; All protocols are per-well of a six-well plate. Hunt1 Margaret J. Reporter gene expression for monitoring gene transfer. The mixture was incubated for 5 min.