I transfected GFP into cells using Lipofectamine 2000 and saw a light granular orange background fluorescence. What could be causing this?
Transfection with cationic lipids can produce light granular orange background fluorescence. The orange fluorescence is associated with the lipid/DNA complexes and is not related to GFP. This background varies depending on the cationic lipid reagent used and does not interfere with transfection results. If desired, try performing fluorescence imaging in PBS instead of culture medium. Also, make sure the cells are healthy and intact as lysed cells or cells under stress could generate autofluorescence products.
Can I use Lipofectamine 2000 to co-transfect plasmids and siRNA?
You can co-transfect plasmids and siRNA using Lipofectamine 2000. Please click here (http://www.thermofisher.com/us/en/home/references/protocols/cell-culture/transfection-protocol/plasmid-co-transfection-protocol-lipofectamine-sirna.html) for protocol information.
For how long is the Lipofectamine 2000:DNA complex stable?
Lipofectamine 2000:DNA complexes are stable for 6 hours at room temperature when the complex is prepared in Opti-MEM medium. Use of other media for complex formation may decrease stability.
Low-density Lipoprotein Receptor-related Protein-1 (LRP1) Mediates Autophagy and Apoptosis Caused by Helicobacter pylori VacA.
Authors:Yahiro K, Satoh M, Nakano M, Hisatsune J, Isomoto H, Sap J, Suzuki H, Nomura F, Noda M, Moss J, Hirayama T,
Journal:J Biol Chem
PubMed ID:22822085
In Helicobacter pylori infection, vacuolating cytotoxin (VacA)-induced mitochondrial damage leading to apoptosis is believed to be a major cause of cell death. It has also been proposed that VacA-induced autophagy serves as a host mechanism to limit toxin-induced cellular damage. Apoptosis and autophagy are two dynamic and opposing processes that ... More