Chloroquine transfection protocol

Discussion in 'Hydroxychloroquine Sulfate 200 Mg' started by vasa666, 10-Mar-2020.

  1. dennis_ru Guest

    Chloroquine transfection protocol


    This procedure can be modified for alternative packaging cell lines or transfection reagents. Once produced, lentivirus can be used for a variety of downstream applications such as stable-cell line generation.

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    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. 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. The following are the protocols I've gathered *4-6 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.

    Test a variety of brands and lots of FBS to find one suitable with your protocols. Last Upload: August 26, 2016 *Pro-Tip* Different brands and lots of FBS can promote or inhibit transfection.

    Chloroquine transfection protocol

    Improve Lentiviral Production Using Lipofectamine 3000., Addgene Lentivirus Production Protocol

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  5. May 07, 2001 Transfection efficiency in fibroblasts showed a linear increase 3–6-fold as the addition of chloroquine increased from 10 to 100 μM compared to plasmid alone Fig. 2 a–c. The maximum gene expression twofold increase compared to lipofectamine alone was observed in CHO cells when the cells were transfected with lipofectamine in the presence of chloroquine Fig. 2 b.

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    The initial transfection is further improved by use of chloroquine. Stocks of ecotropic virus, collected as culture supernatant 48 hrs. post-transfection, can be stored at -80°C and used for infection of cell-lines in view of transformation and in vitro studies, or primary cells such as mouse bone marrow cells, that can then be used for transplant in our mouse model. I have different protocols for Lentivirus production using Lipofectamine transfection of HEK293T cells and now am confused about the concentration of Chloroquine and Na-Butyrate. 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.

     
  6. bally Well-Known Member

    Plaquenil (Hydroxychloroquine sulfate) is a drug originally used to prevent or treat malaria. Hydroxychloroquine Plaquenil Side Effects & Dosage for Malaria Plaquenil Uses, Dosage & Side Effects - Plaquenil Side Effects on Your Eyes and Vision
     
  7. otaha User

    If you subscribe to any of our print newsletters and have never activated your online account, please activate your account below for online access. First time on Plaquenil - Side Effects of Plaquenil Hydroxychloroquine, Warnings, Uses Plaquenil Hydroxychloroquine - Side Effects, Dosage.
     
  8. Iohanan Well-Known Member

    Hydroxychloroquine Plaquenil Hydroxychloroquine Plaquenil is considered a disease-modifying anti-rheumatic drug DMARD. It can decrease the pain and swelling of arthritis. It may prevent joint damage and reduce the risk of long-term disability. Hydroxychloroquine is in a class of medications that was first used to prevent and treat malaria.

    Hydroxychloroquine Oral Uses, Side Effects, Interactions.
     
  9. de facto Well-Known Member

    This is Why You Need Multifocal ERG if You Have Plaquenil. Testing for Chloroquine and Hydroxychloroquine Retinopathy. Introduction Although the exact mechanism by which chloroquine CQ and hydroxychloroquine HCQ - commonly known as Plaquenil induce retinal toxicity is not well understood, but it is recognized as a serious ophthalmologic concern because it is not treatable. 1 Guidelines from the American Academy of Ophthalmology AAO on screening.

    Despite Plaquenil dosing recommendations, retinal toxicity.