Alternatively, we suggest using this opportunity to take a little break from work and read some of the interesting articles below. This protocol describes a general method for transfecting mammalian cells using linear polyethylenimine. Plaquenil treatment rheumatoid arthritis Plaquenil addictive Reactions to plaquenil Chloroquine cryptocaryon The plasmid DNA pGFP-N2 0.2μg and transfection reagents Lipofectamine 2000 were diluted in Opti-MEM 25μl, and then the diluted transfection reagent dropping 25μl dilutions of plasmid DNA to 25μl, the side with the vortex oscillator plus side, at room temperature and incubated for 20 min. The weight Transfection efficiency was affected by the concentration of lysosomotropic agents. The marked transfection enhancement was observed when the amount of sucrose increased from 5 to 500 mM and the optimum transfection efficiency was found at 500 mM in all cell lines tested Fig. 2 a–c. Transgene expression was found to be very low when the cells were transfected with plasmid alone. Hours after transfection reaction has been applied to cells, gently remove media and replace with 10% glycerol or DMSO for 2-3 minutes. Remove and feed with complete media various references. *1 hour before transfection, replace cDMEM with cDMEM + 25 mM chloroquine. Add transfection reaction to this media, and incubate o/n JoVE. We specifically use this protocol with Lenti-X 293T cells, a cell line optimized for production of lentiviral vectors. Transfections allow for transient expression of a gene of interest in a target cell line and can be useful for short term studies of protein function. Chloroquine transfection lipofectamine High efficiency polyoma DNA transfection of chloroquine treated cells., Enhanced plasmid DNA transfection with lysosomotropic. Dmards plaquenil methotrexatePlaquenil and diarrhea eventually goea awayHydroxychloroquine sulfate for hair loss The intracellular trafficking mechanism of Lipofectamine-based transfection reagents and its implication for gene delivery. Skip to main content. Thank you for visiting. The intracellular trafficking mechanism of Lipofectamine-based.. Glycerol shock or chloroquine? - Transfection and Transduction. Transient Transfection of 293T cells. Transfection efficiency was affected by the concentration of lysosomotropic agents. The marked transfection enhancement was observed when the amount of sucrose increased from 5 to 500 mM and the optimum transfection efficiency was found at 500 mM in all cell lines tested Fig. 2 a–c. Chloroquine diphosphate 52 mg Deionized, distilled H 2 O 1 mL Dissolve 52 mg of chloroquine diphosphate in 1 mL of deionized, distilled H 2 O. Sterilize the solution by passing it through a 0.22-µm filter. Store the filtrate in foil-wrapped tubes at −20°C. Transfection efficiency was significantly increased by the treatment of chloroquine, indicating that endocytosis possibly was the major cellular uptake pathway. Compared to Lipofectamine™ system, this new delivery system demonstrated similar transfection efficiency but a much lower cytotoxicity.