Chloroquine transfection protocol

Discussion in 'Canadian Pharmacy Online' started by Grummer, 10-Mar-2020.

  1. DARKNESS2007 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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    Our laboratory uses PEI over other cell transfection reagents because of its low cost. This protocol is appropriate for two suspension cell lines, CHO-S and HEK 293 GnTi-. Many cell lines can be transfected successfully with PEI but in our experience these two cell lines express the highest level of protein compared to other cells. Prior to adding complex, remove 1 mL of medium from each well, leaving a total of 1 mL in each well. Add 500 μL of lipid–DNA complex to each well, taking care to dispense liquid against the well wall to avoid disrupting cells. Gently agitate plate to evenly distribute. Incubate plate for 6 hours at 37°C, 5% CO 2. The following protocol is for 10 cm dish medium 10ml. 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. 1. Plate cells the night before to give 60-70% confluence at the day of transfection.

    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

    Troubleshooting Transfection Experiments Thermo Fisher., Improve Lentiviral Production Using Lipofectamine 3000.

  2. Neostigmine chloroquine phosphate
  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.

    • Addgene Lentivirus Production Protocol.
    • Transient transfection into 293T cells by Jun Takagi, Jun 15..
    • Questions with answers in CHLOROQUINE Science topic.

    Day 2 Phoenix cell transfection & target cell preparation Part 1 Phoenix cell transfection 1. Feed Phoenix cells with DMEM supplemented with chloroquine diluted from 1000x stock to 1x 30 min before transfection. 2. In a FACS tube in the hood, add the following 15 µg DNA 40 µL 2M CaCl 2 MilliQ water to final volume of 500 µL 3. This protocol is a modified version of a published method 4, in which calcium phosphate–based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney. Since the publication of the original method1, increases in efficiency have been achieved by including additional steps such as glycerol shock2 and/or chloroquine treatment3. This protocol is a modified version of a published method4, in which calcium phosphate–based transfection methods for Chinese hamster ovary cells and the 293 line of human embryonic kidney cells were rigorously optimized.

     
  4. BASM Well-Known Member

    Chloroquine is used to prevent or treat malaria caused by mosquito bites in countries where malaria is common. CBD Oil and Medical Marijuana For Arthritis & RA Joint Pain Chloroquine Oral Route Proper Use - Mayo Clinic Chloroquine Oral Uses, Side Effects, Interactions, Pictures, Warnings.
     
  5. nnt 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 Indications, Side Effects, Warnings - Chloroquine Side Effects Common, Severe, Long Term - Chloroquine - Wikipedia
     
  6. creol Guest

    Hydroxychloroquine Plaquenil Toxicity and Recommendations. Chloroquine CQ and hydroxychloroquine Plaquenil HCQ have been used for many years, initially for the treatment of malaria but now more commonly for the treatment of inflammatory diseases such as rheumatoid arthritis and lupus 1. It is now considered for new disease applications in diabetes, heart disease and adjunct cancer therapies 2.

    Eye screening for patients taking hydroxychloroquine.