Chloroquine transfection calcium phosphate

Discussion in 'Plaquenil' started by b3dun, 24-Feb-2020.

  1. yarax New Member

    Chloroquine transfection calcium phosphate


    If you use 6-well or 12-well plates, total volume of the medium should be 3ml and 1.5ml, respectively, and decrease the amount of each reagent accordingly.2. One hour prior to the transfection, change to medium containing 25M chloroquine (from x1000 stock in PBS, stored at -20C). (Chloroquine can be omitted, but increases efficiency about x2) to a final volume of 500 l.

    Lysosomal inhibitor chloroquine 2016 ophthalmology journal hydroxychloroquine Discoid lupus plaquenil

    Chloroquine diphosphate CQ was ingeniously used to take place of phosphate salt in traditional calcium phosphate coprecipitation method for pDNA transfection. With multiple roles of CQ in the novel Ca-CQ-pDNA complex including pDNA compaction and assistance in lysosome escape, the transfection efficiency of the pDNA was significantly increased relative to the traditional method. CQ did not. The optimized transfection procedure requires that CHO cells to be incubated with calcium phosphate-DNA coprecipitate and chloroquine 100 microM for a total of 16 h. By using high-molecular-weight human genomic DNA as a DNA source for transfection, we show that this procedure is equally efficient for stably transferring a much larger gene. Transfection of 293T cells • Calcium Phosphate transfection reagents self-made or kits, e.g. Promega ProFection or Sigma #CAPHOS. • 25mM Chloroquine in PBS.

    Add this mixture directly to the cells dropwise through the medium. As long as the cells are alive, they produce proteins. When you are ready, add 500 l of 2x HBS dropwise while gently mixing. Make sure you evenly sprinkle the droplet over the entire area.5. For secreted protein, you can change media once (at day 3 or 4, save the collected sup) and give it another 3-4 days of culture.

    Chloroquine transfection calcium phosphate

    Transfection - Wikipedia, High-efficiency stable gene transfection using chloroquine-treated.

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  7. Chloroquine treatment of rodent cells during the first hours of polyoma DNA transfection increase the fraction of cells expressing viral functions. The effect has been observed after DNA absorption using both the DEAE-dextran and calcium phosphate coprecipitation methods.

    • High efficiency polyoma DNA transfection of chloroquine treated cells..
    • Transfection of 293T cells - Kristoffer Riecken.
    • Plasmid transfection in HEK293T cell by Calsium Phosphate..

    I have another question regarding calcium phosphate transfections. I am planning to do another this week, but I would like to compare standard CaPO4 transfection efficiencies w/those of modified techniques. I've read several articles suggesting a glycerol shock and/or adding chloroquine to the media. Phosphate solution, and allowing the mixture to incubate at room temperature. This step generates a precipitate that is dispersed onto the cultured cells. The precipitate is taken up by the cells via endocytosis or phagocytosis. Reagent-Based Methods Calcium Phosphate Ca++ Ca++ Ca++ Ca++ Calcium phosphate Ca++ Ca++ DNA Common Transfection. Add 22 l of 25mM chloroquine to each 15cm dish for 10cm plate 7.8 l of chloroquinei.e. final concentration = 25 M. The cells should be transfected within 5-10’ of adding the chloroquine. 7. For 15cm plates, add 1.69ml of the transfection buffer to the DNA-CaCl2 solution by “bubbling”.

     
  8. zmeuka New Member

    Plaquenil (hydroxychloroquine) belongs to a group of medicines called quinolines. Plaquenil - FDA prescribing information, side effects and uses The Risks of Pregnancy for Women With Lupus - Lupus Center. Treating Lupus with Anti-Malarial Drugs Johns Hopkins.
     
  9. KAmaR Guest

    Hydroxychloroquine sulfate and Methotrexate drug interactions. This study is created by eHealthMe based on reports of 5,663 people who take Hydroxychloroquine sulfate and Methotrexate from Food and Drug Administration FDA, and is updated regularly. On eHealthMe, you can evaluate side effects and drug interactions from 16 million FDA reports. The results are personalized to your gender and age 0-99+.

    Methotrexate and hydroxychloroquine treatment Rheumatoid.
     
  10. Axeomica XenForo Moderator

    Chloroquine Phosphate - WebMD Chloroquine Phosphate. Chloroquine is used to prevent or treat malaria caused by mosquito bites in countries where malaria is common. Malaria parasites can enter the body through these mosquito bites, and then live in body tissues such as red blood cells or the liver.

    Chloroquine Phosphate Chloroquine Hydrochloride Injection