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. We specifically use this protocol with Lenti-X 293T cells, a cell line optimized for production of lentiviral vectors. Chloroquin site drugs.com Synthroid and plaquenil Who makes generic plaquenil Transfection Procedure. The morning of the transfection day, replace the media with fresh DMEM without PSG and containing 10 uL of 25 mM chloroquine optional. Wait ~5h before going onto the next step. Prepare DNA in a sterile 1.5 mL tube or tubes as needed. Use 100 uL per well. Add 3 uL PEI per ug of DNA in OptiMEM. Add DNA/CaCl2-solution to 2x HBS drop wise, while blowing air through HBS with pasteur pipette. • Incubate the mixture for 10 to 20min at room temperature. • Remove old medium from the cells. • Add 10ml medium including 25µM Chloroquine the Chloroquine is used in this step only. A pH-responsive peptide fragment modelled on the influenza virus haemagglutinin INF7-SGSC can promote the transfectional activity of polyL-lysine pLL/DNA complexes against 293 cells. Last Upload: June 10, 2016 Day 0: Seed Lenti-X 293T cells (this cell line is optimized for production of lentiviral vectors) Day 1 (pm): Transfect Cells Day 2 (am): 18h post transfection - Remove media, replace with fresh media Day 3 or more (am): Observe fluorescence, harvest cells, or perform your experiment *Pro-Tips* Different brands and lots of FBS can promote or inhibit transfection. This approach can be adapted for different cell lines and different transfection reagents. Chloroquine dna transfection Transfection Guide Overview of Transfection Methods Promega, Transfection of 293T cells - Kristoffer Riecken Chloroquine and paludrine in pregnancyChloroquine or mefloquine resistance malaria Chloroquine is a 4-aminoquinoline with antimalarial, anti-inflammatory, and potential chemosensitization and radiosensitization activities. Although the mechanism is not well understood, chloroquine is shown to inhibit the parasitic enzyme heme polymerase that converts the toxic heme into non-toxic hemazoin, thereby resulting in the accumulation of toxic heme within the parasite. Chloroquine C18H26ClN3 - PubChem. Chloroquine and amphipathic peptide helices show synergistic.. Transfection Methods Overview - Bio-Rad. Prepare the DNA in HBS for application to cells. 1. About 5 minutes prior to transfection, add chloroquine to each plate to 25uM chloroquine stock is 50 mM; for 3 mL media + 1 mL DNA, add 2u1. Chloroquine acts to inhibit lysosomal DNases by neutralizing vesicle pH. Chloroquine is a member of quinolone family and is a weak intercalating agent. Chloroquine is used for treating amebiasis, rheumatoid arthritis, discoid and systemic lupus erythematosus. Application DNA intercalator. Also used to increase transfection efficiency. Chloroquine diphosphate salt has been used • in in vitro antiplasmodial assays The chloroquine enhanced transfection procedure provides a reliable and reproducible assay of released protein and its effect is presumably mediated through its pH elevating effect on the lysosomal compartment, thereby, protecting DNA molecules from nuclease degradation.