Most non-viral vectors are known to internalize in the cells by endocytosis. Therefore, low transfection efficiency of non-viral vectors may be due to intracellular degradation of input DNA in the endosomes and/or lysosomes. Plaquenil moa Chloroquine 500mg tab Does plaquenil reduce rheumatoid arthritis Hydroxychloroquine foods to avoid Chloroquine is an inhibitor of the lysosomal degradation of the DNA which is taken up by the cells, so as leelee said, transfection should have been successful, albeit at a slightly lower level than if you would have added the chloroquine. In my normal transfections, I never add chloroquine and get high efficiencies nevertheless. 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. Carefully transfer the transfection mix to the Lenti-X 293T packaging cells. Add the transfection mix dropwise being careful not to dislodge the cells. Incubate the cells for 18 h, or until the following morning. The following morning, carefully aspirate the media. Replace the media with 15 mL of DMEM complete. We report here the effects of individual lysosomotropic agents such as chloroquine, polyvinylpyrolidone (PVP) and sucrose on β-gal expression in cultured fibroblasts COS, 293 and CHO. DNA degradation can be inhibited either by inactivating the lysosomal enzymes or obliterating endosome fusion to lysosomes using lysosomotropic agents. Chloroquine concentration transfection Chloroquine Diphosphate 100 mm for Transfection, Helper Dependent Protocol - Stanford University Is plaquenil a steroid an immunosuppressant Chloroquine stocks 25 mM stocks. See Preparation of Chloroquine Stocks. PEI 1 mg/mL stocks. See Preparation of PEI Stocks. DMEM/10% FBS/PSG and DMEM/10% FBS with no PSG; 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. PEI Mediated Plasmid Transfection - Bridges Lab Protocols. Addgene General Transfection. Chloroquine C18H26ClN3 - PubChem. If kept for extended periods of time, chloroquine is toxic. The solution in the plates looks fuzzy, due to a very fine, dust-like precipitation of the transfection mixture. Note It is important to do all media changes with extreme gentleness, as the cells have been sensitized by the chloroquine, and can easily detach from the plate. The concentration of chloroquine added to the growth medium and the duration of treatment are limited by the sensitivity of the cells to the toxic effect of the drug and should be determined. Figure 4. Transfection optimization using the ViaFect™ Transfection Reagent. TF-1 cells were plated in growth media without antibiotics at 30,000 cells per well in a white 96-well assay plate and transfected with a CMV-luc2 plasmid using various lipid ViaFect™ Transfection ReagentDNA ratios. The DNA concentration was held constant at.