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 retinopathy vs diabetic retinopathy Hydroxychloroquine interaction with cyclobenzaprine Add 22 l of 25mM chloroquine to each 15cm dish for 10cm plate 7.8 l of chloroquinei.e. final concentration = 25 M. The cells should be transfected within 5-10’ of adding the chloroquine. 7. For 15cm plates, add 1.69ml of the transfection buffer to the DNA-CaCl2 solution by “bubbling”. Excretion of Chloroquine is quite slow,but is increased by acidification of the urine. Chloroquine is deposited in the tissues in considerable amounts. In animals, from 200 to 700 times the plasma concentration may be found in the liver,spleen, kidney, and lung; leukocytes also concentrate the drug. 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. 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 Questions with answers in CHLOROQUINE Science topic, Chloroquine - FDA prescribing information, side effects and uses Chloroquine free baseInfo plaquenil To understand how chloroquine CQ enhances transgene expression in polycation-based, nonviral gene delivery systems, a number of CQ analogues with variations in the aliphatic amino side chain or in the aromatic ring are synthesized and investigated. Our studies indicate that the aliphatic amino moiety of CQ is essential to provide increased gene expression. Structure−Function Correlation of Chloroquine and.. Calcium phosphate–mediated transfection of eukaryotic cells.. Addgene General Transfection. 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. Chloroquine enters cells and accumulates in vesicular compartments; the overall intracellular concentration increases to 9 mM,which means that in the vesicular compartment, the chloroquine concentration is still higher. At such high concentrations, chloroquine induces the dissociation of plasmid DNA/lactosylated polylysine complexes, as shown. 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.