Chloroquine in transfection

Discussion in 'Generic & Brand Canadian Pharmacy' started by .trixs, 18-Mar-2020.

  1. wineart Well-Known Member

    Chloroquine in 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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    Chloroquine is a member of quinolone family and is a weak intercalating agent. Chloroquine is used for treating amebiasis, rheumatoid arthritis, discoid and systemic lupus erythematosus. Application DNA intercalator. Also used to increase transfection efficiency. Chloroquine diphosphate salt has been used • in in vitro antiplasmodial assays 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. So we use Chloroquine as a transfection agent at the concentration 25uM for 8 hours. Than changing the media. So far we detected much higher transfection efficiency with Chloroquine than.

    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 in transfection

    Helper Dependent Protocol - Stanford University, Glycerol shock or chloroquine? - Transfection and Transduction

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  4. Chloroquine has been frequently used in cell culture gene transfection studies Cotton et al. 1992, Wagner et al. 1992, Lucas et al. 1995, but has not been reported yet to be successful for enabling gene transfection in vivo. As mentioned above, the mechanisms of action of chloroquine seems to be related to elevating the intravesicular pH of lysosomes and endosomes, thereby providing a suboptimal pH environment for the various lysosomal enzymes including those involved in DNA degradation.

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    May 07, 2001 Chloroquine has been frequently used in cell culture gene transfection studies Cotton et al. 1992, Wagner et al. 1992, Lucas et al. 1995, but has not been reported yet to be successful for enabling gene transfection in vivo. As mentioned above, the mechanisms of action of chloroquine seems to be related to elevating the intravesicular pH of lysosomes and endosomes, thereby providing a suboptimal pH environment for the various lysosomal enzymes including those involved in DNA degradation. 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. Chloroquine is an aminoquinoline that is quinoline which is substituted at position 4 by a 5-diethylaminopentan-2-ylamino group at at position 7 by chlorine. It is used for the treatment of malaria, hepatic amoebiasis, lupus erythematosus, light-sensitive skin eruptions, and rheumatoid arthritis.

     
  5. GORKOM New Member

    In some cases, they may not be available in every strength or form as the brand-name drug. Three Reasons Why Your Thyroid Medication Isn't Working Chris Kresser Plaquenil Uses, Dosage & Side Effects - Plaquenil Hydroxychloroquine for Lupus - LupusCorner
     
  6. maximixer Well-Known Member

    In deciding to use a medicine, the risks of taking the medicine must be weighed against the good it will do. For this medicine, the following should be considered: Tell your doctor if you have ever had any unusual or allergic reaction to this medicine or any other medicines. Plaquenil allergy signs - Answers on HealthTap Plaquenil hydroxychloroquine 200mg is it causing itchy. Allergic reaction to Plaquenil Lupus Forums at The Lupus.
     
  7. SI2 Moderator

    Preventive Approach to Congenital Heart Block With Hydroxychloroquine. Preventive Approach to Congenital Heart Block With Hydroxychloroquine PATCH The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U. S. Federal Government.

    Hydroxychloroquine in lupus pregnancy - Clowse - 2006 - Arthritis &.