Chloroquine retroviral transfection

Discussion in 'Hydroxychloroquine' started by Homyak, 12-Mar-2020.

  1. Madmax_s XenForo Moderator

    Chloroquine retroviral 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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    Retrovirus Transfection Protocol Prepared by Berggren, Travis [email protected] Submitted April 24, 2012 Submitted by Berggren, Travis [email protected] from Salk Stem Cell Core in-house protocols Contributors Lutz, Margaret. Modesto, Veronica. Panopoulos, Athanasia. Affiliation The Salk Institute Introduction In this study, the effect of chloroquine on transduction and associated toxicity of baboon CD34 cells and a variety of cell lines was studied. Materials and Methods. Cell lines. To study the effects of chloroquine on retroviral transduction of cell lines, three cell lines were chosen Hela cells, D17 and 208 F cells. Here, we co-transfect 293 cells with a retroviral vector expressing the oncogene of interest and a plasmid that expresses the gag-pol-env packaging functions, such as the single-genome packaging constructs kat or pCL, in this case the EcoPak plasmid. The initial transfection is further improved by use of chloroquine.

    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 retroviral transfection

    Principles and Mechanisms of Mammalian Cell Transfection., EFFECT OF CHLOROQUINE ON TRANSDUCTION OF CELL LINES AND.

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  5. Ing for transfection. In general, the cells should be plated at a density so that they are 95-100% confluent at 24 hours post-transfection. * The addition of chloroquine to the medium appears to increase retroviral titer by approximately two fold. This effect is presumably due to the lysosomal neutralizing activity of the chloroquine.

    • PhoenixTM Retroviral Packaging Cell Lines Ampho and Eco.
    • Production of Replication-Defective Retrovirus by Transient..
    • Addgene General Transfection.

    Chloroquine diphosphate salt C6628 - Product Information Sheet Author Sigma-Aldrich Corp. Subject This product is a member of the quinoline family with multiple applications. It was originally used as an antimalarial compound. Retroviral calcium phosphate transfection - How much DNA should I use. does anyone have anyone have any experience using chloroquine or a glycerol shock in transfections? Any input is greatly appreciated. Thanks so much! P. S. Any transfection tips for A293T or Phoenix cell lines, using CaPO4, Mirus, lipofectamine, or whatever, are warmly. MM chloroquine 1000x Sigma C6628 8 mg/mL polybrene/hexadimethrine bromide 1000x Sigma H9268 2M CaCl 2 2x HBS see recipe at end of protocol DNA retroviral vector Procedure Day 1 Phoenix cell preparation 1. Plate cells at 2.5×106 cells in 8 mL DMEM in a 10 cm dish one for each transfection to be done. System

     
  6. Gracie User

    At Healthfully, we strive to deliver objective content that is accurate and up-to-date. Chagas Disease Symptoms & 6 Natural Treatments - David. Treatment of Chagas - Chagas Disease Center of Excellence COULD I HAVE CHAGAS DISEASE? - CHAGAS COALITION
     
  7. neocpeter Guest

    We use cookies to make interactions with our website easy and meaningful, to better understand the use of our services, and to tailor advertising. Lysosomal membrane permeabilization and cell death. Abstract 3511 Chloroquine-induced lysosomal membrane permeabilization. Lysosomotropic drugs activate TFEB via lysosomal membrane fluidization.
     
  8. gnikolay New Member

    Is it safe to take plaquenil hydroxychloroquine while pregnant. Dr. Anzalone responded Not safe. "usage of this drug during _hyperlink_clicked"pregnancy/a should be avoided except when in the judgment of the physician the benefit outweighs the possible hazard.

    Hydroxychloroquine - Wikipedia