Chloroquine in transfection

Discussion in 'Aralen 250 Mg' started by zhuk, 04-Mar-2020.

  1. Smersh Moderator

    Chloroquine in transfection


    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.

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    PKa 8.4 and 10.8 for chloroquine base at 20 °C 2. This product is a member of the quinoline family with multiple applications. It was originally used as an antimalarial compound.3 It kills the erythrocytic forms of malaria parasites at all stages of development, but does not affect the sporozoites. Transfection Methods Reagent-Based Methods DEAE-Dextran Method Overview Solution A DNA ~1–5 µg/ml diluted into 2 ml of growth medium with serum containing chloroquine Solution B DEAE-dextran solution ~50–500 µg/ml Solution C ~5 ml of DMSO Solution D Complete growth medium 1 Add solution A to solution B, then mix gently. Test a variety of brands and lots of FBS to find one suitable with your protocols. FBS can be purchased already heat inactivated or it can be inactivated in the lab by heating to 56 ℃ for 30 minutes. 25 mM chloroquine diphosphate Dissolve 0.129 g of chloroquine diphosphate salt in 10 mL of sterile water.

    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 in transfection

    Addgene General Transfection, Transfection Methods Overview - Bio-Rad

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  7. Application. DNA intercalator. Also used to increase transfection efficiency. Chloroquine diphosphate salt has been used • in in vitro antiplasmodial assays • in transfection and infection assays • in autophagy inhibition • in differentiation of induced pluripotent stem iPS cells into cardiomyocytes • in flow treatment of infected blood.

    • Chloroquine diphosphate salt powder or crystals, 98.5-101.0%..
    • Addgene Lentivirus Production Protocol.
    • Putative Role of Chloroquine in Gene Transfer into a Human..

    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. Chloroquine-resistant cells efflux chloroquine at 40 times the rate of chloroquine-sensitive cells; the related mutations trace back to transmembrane proteins of the digestive vacuole, including sets of critical mutations in the P. falciparum chloroquine resistance transporter PfCRT gene. One hour prior to the transfection, change to medium containing 25µM chloroquine from x1000 stock in PBS, stored at -20°C. Volume should be 10 ml per dish. Chloroquine can be omitted, but increases efficiency about x2 3. Add 10µg DNA and ddH2O to 438 µl in 15-ml sterile tube.

     
  8. dji Guest

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  9. Grom Well-Known Member

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  10. FireBear Well-Known Member

    Stopping Plaquenil - General Discussion - Life With Lupus So I ran out of my plaquenil and the Rheum. that prescribed it I am no longer seeing and wont re-fill it and I dont see my new Rheum. until Decemeber 23rd. I am nervous to see what will happen. The pharmacist said there wont be any danger just stopping it. Im just nervous to see what my body will do. Has anyone gone off the plaquenil and what happened? I hear my new Rheum. is all for it so I.

    Does anyone know if you have to be weaned off Plaquenil? I.