Chloroquine transfection

Discussion in 'Canadian Pharmacy Online' started by Sid115, 19-Mar-2020.

  1. Zilot Guest

    Chloroquine transfection


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    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. Chloroquine is a lysosomotropic agent that prevents endosomal acidification 1. It accumulates inside the acidic parts of the cell, including endosomes and lysosomes. This accumulation leads to inhibition of lysosomal enzymes that require an acidic pH, and prevents fusion of endosomes and lysosomes. Transfection of 293T cells • Calcium Phosphate transfection reagents self-made or kits, e.g. Promega ProFection or Sigma #CAPHOS. • 25mM Chloroquine in PBS.

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    Chloroquine transfection

    EFFECT OF CHLOROQUINE ON TRANSDUCTION OF CELL LINES AND., Chloroquine for research Cell-culture tested InvivoGen

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  4. 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.

    • Transfection Methods Overview - Bio-Rad.
    • Transfection of 293T cells - Kristoffer Riecken.
    • 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. Plate cells the night before to give 60-80% confluence at the day of transfection. 2. 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. Carefully transfer the transfection mix to the Lenti-X 293T packaging cells. Add the transfection mix dropwise being careful not to dislodge the cells. Incubate the cells for 18 h, or until the following morning. The following morning, carefully aspirate the media. Replace the media with 15 mL of DMEM complete.

     
  5. twobarrels_shadow Well-Known Member

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  6. lotar Moderator

    Hydroxychloroquine is widely used in the treatment of post-Lyme arthritis. Plaquenil caused me to go into menopause lupus Plaquenil Side Effects Common, Severe, Long Term - Common and Rare Side Effects for Plaquenil Oral
     
  7. probiz User

    Plaquenil hydroxychloroquine sulfate dosing, indications. Medscape - Indication-specific dosing for Plaquenil hydroxychloroquine sulfate, frequency-based adverse effects, comprehensive interactions, contraindications, pregnancy & lactation schedules, and cost information.

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  8. Gravi New Member

    Hydroxychloroquine Side Effects, Dosage, Uses, and More Hydroxychloroquine Plaquenil is used to treat malaria, lupus, and rheumatoid arthritis. Learn about side effects, warnings, dosage, and more.

    Plaquenil Hydroxychloroquine - Side Effects, Dosage.