Chloroquine pretreatment before transfection

Discussion in 'Canadian Pharmacy Drugs Online' started by Levkaink, 11-Mar-2020.

  1. MVK2 New Member

    Chloroquine pretreatment before 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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    Both adults and children should take one dose of chloroquine per week starting at least 1 week before. traveling to the area where malaria transmission occurs. They should take one dose per week while there, and for 4 consecutive weeks after leaving. The weekly dosage for adults is 300mg base 500mg salt. Autophagy inhibition through chloroquine pretreatment or Atg5shRNA infection led to the increase of cell apoptosis after PA treatment. Moreover, induction of autophagy by pretreatment with rapamycin resulted in distinct decrease of PA-induced apoptosis. In the current study, we observed that pretreatment of COS7 and HEK293 cells with these lysosomotropic agents, chloroquine 100 μM and ammonium chloride 10 mM, for 4 hours prior to electrotransfection insignificantly altered eTE Fig 7a and 7b. The pretreatment was required because the lysosomotropic agents need to be in lysosomes before.

    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 pretreatment before transfection

    Chloroquine ameliorates carbon tetrachloride-induced acute., Autophagy protects against palmitate-induced apoptosis in hepatocytes.

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  7. The optimal transfection time depends on the cell line, transfection reagent and nucleic acid used. For the ViaFect™ Transfection Reagent and the FuGENE® HD Transfection Reagent, which are two of the more gentle methods of DNA transfection into cells, there is no need to add more medium or replace the medium after transfection.

    • Transfection Guide Overview of Transfection Methods Promega.
    • Distinct effects of endosomal escape and inhibition of..
    • Transient Transfection of 293T cells.

    Chloroquine has been used to increase transfection efficiency for calcium phosphate, DEAE-dextran, and some liposome-mediated gene delivery 63,65–68. However, effects of lysosomotropic agents on polymer and some lipid based methods of gene delivery are still controversial in the literature. Chloroquine This drug is taken once a week, starting about 1 to 2 weeks before your trip and continuing for 4 weeks after. But chloroquine is rarely used anymore, because it no longer works. Pretreatment with 0.1, 1, and 10 μM chloroquine reduced infectivity by 28%, 53%, and 100%, respectively. Reproducible results were obtained from three independent experiments. Reproducible results were obtained from three independent experiments.

     
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