What is Lipofectamine LTX?
Lipofectamine® LTX Reagent is a plasmid transfection reagent that offers the right balance of potency and gentleness for your cells, resulting in the highest possible transfection efficiencies and viabilities.
What is lipofectamine transfection?
Lipofectamine or Lipofectamine 2000 is a common transfection reagent, produced and sold by Invitrogen, used in molecular and cellular biology. It is used to increase the transfection efficiency of RNA (including mRNA and siRNA) or plasmid DNA into in vitro cell cultures by lipofection.
What is liposome mediated transfection?
Using liposomes to deliver DNA into different eukaryotic cell types results in higher efficiency and greater reproducibility than other transfection methods. For transient expression of DNA in the nucleus, plasmid DNA is complexed with a liposome suspension in serum-free medium.
How do liposomal transfection reagents work?
Liposome transfection (i.e. lipofection) is a technique that results in the transfer of genetic material into a cell by means of liposomes. Lipid based reagents use cell membrane semi-permeability to deliver foreign nucleic acid molecules into a cell.
How do you transfect HEK293 cells?
The protocol for a 24-well transfection reaction with HEK293 cells is here:
- Plate 10,000-15,000 HEK293 cells per well in 0.5 ml of complete growth medium 12-24 hours prior to transfection.
- Wash with 1xPBS and add 0.5 ml of fresh growth medium.
What is Lipofectamine Plus reagent?
Lipofectamine LTX Reagent is a plasmid transfection reagent that offers a balance of potency and gentleness for your cells, resulting in high transfection efficiencies and viabilities.
Is Lipofectamine stable transfection?
Stable transfection requires the transfected DNA to integrate itno the cell’s chromosomes. I do stable transfections with Lipofectamine, so that is not a problem. It has been said that linearized plasmids integrate better than supercoiled plasmids, but supercoiled plasmids give better rates of transfection.
What are the steps of transfection?
- encapsulation of genetic material with transfection reagent.
- Cellular uptake of nanoparticles.
- Release into the cytosol and if needed transport into the nucleus for transcription.
How liposome is used in gene transfer?
Liposomes are often used to deliver a molecular cargo such as DNA for therapeutic benefit. The lipids used to form such lipoplexes can be cationic, anionic, neutral, or a mixture thereof.
Does transfection use lipids to transfer DNA?
Lipofection, also known as “lipid transfection” or “liposome-based transfection,” uses a lipid complex to deliver DNA to cells. Lipids are a broad class of fat-soluble biomolecules, such as fats, oils, and waxes.
How does lipid based transfection work?
Lipofection, also known as “lipid transfection” or “liposome-based transfection,” uses a lipid complex to deliver DNA to cells. Depending on the liposome and cell type, the liposome can be endocytosed (Figure 11.6D) or directly fuse with the cell membrane to release the DNA construct into cells (Figure 11.6E).
What is the best transfection reagent for HEK293?
Conclusion: Lipofectamine 3000 is the best choice for transfection of CHO-K1 and HEK293 with pCDH while Turbofect is preferably used in transfecting these cell lines with pEGFP-N1 (Tab.