Chloroquine concentration transfection

Discussion in 'Canadian Drug' started by okingniels, 27-Feb-2020.

  1. Apocalypse Well-Known Member

    Chloroquine concentration transfection


    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.

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    C for 2 h. Chloroquine was added to each well to a final concentration of 100 µM, and the incubation was con-tinued for an additional 3 h. Medium was aspirated from the wells, and the cells were carefully washed in 1.0 mL of PBS. Wells were replenished with 2 mLof R10 and incubated in 5% CO 2 at 37°C for a total of 24 h from the start of. 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. 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”.

    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

    Optimization of Plasmid DNA Transfection Thermo Fisher., PEI Mediated Plasmid Transfection - Bridges Lab Protocols

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  7. 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.

    • Helper Dependent Protocol - Stanford University.
    • Transient Transfection of 293T cells.
    • The lysosomal inhibitor, chloroquine, increases cell surface..

    Typically, DNA is suspended in sterile water or TE buffer to a final concentration of 0.2–1 mg/ml The optimal amount of DNA to use in the transfection will vary widely depending upon the type of DNA, transfection reagent/method, target cell line, and number of cells Genetic Material DNA Quality and Quantity Factors Affecting A severe eye problem has happened with chloroquine. This may lead to lasting eyesight problems. The risk may be higher if you have some types of eye or kidney problems. The risk may also be higher with some doses of chloroquine, if you use chloroquine for longer than 5 years, or if you take certain other drugs like tamoxifen. 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.

     
  8. Craftworker Moderator

    Hydroxychloroquine is a quinoline medicine used to treat or prevent malaria, a disease caused by parasites that enter the body through the bite of a mosquito. Plaquenil hydroxychloroquine Alternatives & Similar Drugs. Hydroxychloroquine MedlinePlus Drug Information Hydroxychloroquine Oral Uses, Side Effects, Interactions.
     
  9. nipler XenForo Moderator

    Recommendations on Screening for Chloroquine and. Find an Ophthalmologist. Abstract Background The American Academy of Ophthalmology recommendations on screening for chloroquine CQ and hydroxychloroquine HCQ retinopathy are revised in light of new information about the prevalence of toxicity, risk factors, fundus distribution, and effectiveness of screening tools.

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  10. lapotos User

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