Chloroquine dna transfection

Discussion in 'Aralen' started by keo, 04-Mar-2020.

  1. olga™ User

    Chloroquine dna 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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    Hour before transfection, replace cDMEM with cDMEM + 25 mM chloroquine. Add transfection reaction to this media, and incubate o/n JoVE. *After "DNA absorption" 4 h again. replace transfection reaction-containing media with cDMEM + 100 mM chloroquine. Change media after 2-5 hours Luthman & Magnusson, 1983. After cells chloroquine-treated or untreated have been exposed to the calcium phosphate–DNA coprecipitate in growth medium for 2–6 h, remove the medium by aspiration and wash the. Chloroquine diphosphate 52 mg Deionized, distilled H 2 O 1 mL Dissolve 52 mg of chloroquine diphosphate in 1 mL of deionized, distilled H 2 O. Sterilize the solution by passing it through a 0.22-µm filter. Store the filtrate in foil-wrapped tubes at −20°C.

    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 dna transfection

    High efficiency polyoma DNA transfection of chloroquine., Calcium phosphate–mediated transfection of eukaryotic cells.

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  4. Quinacrine QC, a CQ analogue with a fused acridinyl structure that can strongly intercalate DNA, enhances transfection similarly to CQ at a concentration 10 times lower, while N4-4-pyridinyl-N1,N1-diethyl-1,4-pentanediamine CP, a CQ analogue that has a weakly intercalating pyridinyl ring, shows no effect on gene expression.

    • Structure−Function Correlation of Chloroquine and Analogues..
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    The total volume of transfection complex medium, DNA and ViaFect™ Transfection Reagent to add per well of a 96-well plate is 5–10μl. The following protocol is a guideline for transfecting approximately 10–20 wells, depending on the volume of ViaFect™ Transfection ReagentDNA mixture used. For additional wells, scale volumes accordingly. 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 before going onto the next step. Prepare DNA in a sterile 1.5 mL tube or tubes as needed. Use 100 uL per well. Add 3 uL PEI per ug of DNA in OptiMEM. A pH-responsive peptide fragment modelled on the influenza virus haemagglutinin INF7-SGSC can promote the transfectional activity of polyL-lysine pLL/DNA complexes against 293 cells.

     
  5. melan91 New Member

    ydroxychloroquine (HCQ; Plaquenil, Sanofi, Bridgewater, NJ) is an antimalarial agent that is also commonly used as a treatment for a variety of rheumatologic and dermatologic conditions, such as rheumatoid arthritis and systemic lupus erythematosus. Dr. Noskow's Case Report Plaquenil Toxicity - Hollywood Eye. Hydroxychloroquine-related retinal toxicity Rheumatology. New Plaquenil Guidelines -
     
  6. igoria2006 Well-Known Member

    Chloroquine has long been used in the treatment or prevention of malaria from Plasmodium vivax, P. malariae, excluding the malaria parasite Plasmodium falciparum, for it started to develop widespread resistance to it. Chloroquine Dosage Guide with Precautions - Chloroquine Oral Uses, Side Effects, Interactions. Chloroquine - FDA prescribing information, side effects and uses
     
  7. yasha User

    Radiation Oncology/Contraindications - Wikibooks, open books for an. Contraindications to Radiation Therapy. Collagen vascular diseases. It is a commonly held notion that collagen vascular diseases--such as lupus, scleroderma, Sjogren's, etc.--are an absolute contraindication to radiotherapy since the effects on normal tissue would be greater, and even severe.

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