What does Lipofectamine 3000 reagent do?
What does Lipofectamine 3000 reagent do?
Lipofectamine 3000 reagent leverages our most advanced lipid nanoparticle technology to enable superior transfection performance and reproducible results. It delivers exceptional transfection efficiency into the widest range of difficult-to-transfect and common cell types (Figure 1) with improved cell viability.
Do antibiotics affect transfection?
Antibiotics. In general, antibiotics can be present in the medium for transient transfection. However, because cationic lipid reagents increase cell permeability, they may also increase the amount of antibiotics delivered into the cells, resulting in cytotoxicity and lower transfection efficiency.
What is Lipofectamine made of?
Lipofectamine consists of a 3:1 mixture of DOSPA (2,3‐dioleoyloxy‐N‐ [2(sperminecarboxamido)ethyl]‐N,N‐dimethyl‐1‐propaniminium trifluoroacetate) and DOPE, which complexes with negatively charged nucleic acid molecules to allow them to overcome the electrostatic repulsion of the cell membrane.
Does pen strep interfere with transfection?
We have found that antibiotics (pep/strep) do not affect the ability of lipofectamine reagents to transfect cells.
How can I increase my transfection?
Improving the Success of Your Transfection
- Transfect healthy, actively dividing cells at a consistent cell density.
- Transfect using high-quality DNA.
- Optimize the amount of DNA used to transfect cells.
- Optimize the transfection reagent:DNA ratio.
- Optimize cell number per well when transfected.
Can you Trypsinize transfected cells?
Yes, it works fine to trypsinize and re-plate cells 24 hours after siRNA transfection.
What is the lipid in Lipofectamine?
3.1. Lipid-mediated transfection relies on cationic lipids, such as lipofectamine, which facilitate DNA and mRNA delivery into cells. The cationic lipids form a unilamellar liposomal structure with a positive surface charge and fuse with nucleic acids to form a transfection complex.
What is Lipofectamine p3000?
As I understand it, p3000 is a lipid which coats the negatively charged DNA. Lipofectamine 3000, when added to the mixture coats p3000 lipid droplets and confers a positive charge to the outside of the liposhperes. This positive charge allows endocytosis of the lipid droplets into the negatively-charged cell membrane.
Why choose Invitrogen Lipofectamine reagents?
Since launching in 1993, Invitrogen Lipofectamine reagents have become the most referenced transfection reagents with over 50,000 citations to date. You continue to choose these reagents due to their wide acceptance as the best and most reliable available.
How much Lipofectamine 2000 do I need to transfect cells?
To transfect cells in different tissue culture formats, vary the amounts of Lipofectamine 2000, nucleic acid, cells, and medium used in proportion to the relative surface area, as shown in the table. With automated, high-throughput systems, a complexing volume of 50 µl is recommended for transfections in 96-well plates.
What is the difference between FuGene HD and Lipofectamine 3000?
Lipofectamine 3000 reagent provided higher GFP transfection efficiency than Lipofectamine 2000 and FuGENE HD reagents for all five cell lines. Figure 2. The transfection protocol for Lipofectamine 3000 was developed to be easy to use while still ensuring optimum performance and reliability in a wide panel of cell lines.
What is the ratio of DNA to Lipofectamine for plasmid transfection?
Prepare complexes using a DNA (μg) to Lipofectamine 2000 (μl) ratio of 1:2 to 1:3 for most cell lines. Transfect cells at high cell density for high efficiency, high expression levels, and to minimize cytotoxicity. Optimization may be necessary (see “Optimizing plasmid DNA transfection”).
