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    Cyclosporin A (Cyclosporine A)
    Cyclosporin A (Cyclosporine A)

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    This product is for research use only, not for human use. We do not sell to patients.
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    InvivoChem Cat #: V1858
    CAS #: 59865-13-3Purity ≥98%

    Description: Cyclosporin A (Cyclosporine A) is a naturally occuring cyclic polypeptide immunosuppressive agent, which binds to the cyclophilin and then inhibits calcineurin with IC50 of 7 nM in a cell-free assay, it is widely used in organ transplantation to prevent rejection. Cyclosporin A is responsible for the opening of the MPTP (mitochondrial permeability transition pore). When tested with retinal ganglion cells, Cyclosporin A showed a high selectivity for the expression of cyclophilin D and prolonged the cells survival. In human First-Trimester Trophoblast Cells, Cyclosporin A treatment promoted cells growth and invasiveness by inhibiting Ca2+/Calcineurin/NFAT signal. Cyclosporin A treatment promoted the apoptosis of T-cell by upregulating Fas/FasL and caspase activities

    References: Science. 1984 Nov 2;226(4674):544-7; Cell. 1991 Aug 23;66(4):807-15; Biol Reprod. 2008 Jun;78(6):1102-10. 

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    Molecular Weight (MW)1202.61 
    FormulaC62H111N11O12 
    CAS No.59865-13-3 
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 100 mg/mL (83.2 mM)
    Water: <1 mg/mL
    Ethanol: 100 mg/mL (83.2 mM)
    Solubility (In vivo)2% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5mg/mL 
    SynonymsCyclosporine A; Ciclosporin; cyclosporin; cyclosporin A


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    In Vitro

    In vitro activity: Cyclosporin A binds to the cyclophilin (immunophilin) in T cells, forms a Cyclophilin-Cyclosporin A complex which the binds to and inhibits calcineurin. Cyclosporin A inhibits calcineurin with IC50 of 7 nM, then blocks the nuclear translocation of NF-AT. Cyclosporin A also affects mitochondria by preventing the MTP (mitochondrial permeability transition pore) from opening with an IC50 of 39 nM.


    Kinase Assay: Purified bovine brain calcineurin and calmodulin are purchased. Reaction mixtures with purified enzyme contains 100 nM calcineurin, 100 nM calmodulin, and 5 μM 32P-labeled phosphopeptide, in 60 μl (total volume) of assay buffer containing 20 mM Tris (pH 8), 100 mM NaCI, 6 mM MgCl2, 0.5 mM dithiothreitol, 0.1 mg of bovine serum albumin per ml, and either 0.1 mM CaCl2 or 5 mM EGTA. Reaction mixtures with cell lysates contains 20 μl of undiluted lysate, 5 μM 32P-labeled phosphopeptide, and 40 μl of assay buffer. Where indicated, reaction mixtures contains 50 μM peptide 412 or 413 and/or 500 nM okadaic acid, a specific inhibitor of phosphatases 1 and 2A; 500 nM okadaic acid is sufficient for inhibition of Ca2+-independent phosphatases, whereas higher concentrations partially inhibit Ca2+-dependent activity as well. After 15 min at 30°C, reactions are terminated by the addition of 0.5 ml of 100 mM potassium phosphate buffer (pH 7.0) containing 5% trichloroacetic acid. Free inorganic phosphate is isolated by Dowex cation-exchange chromatography and quantitated by scintillation counting as described. 


    Cell Assay: Immunosuppressive agents are dissolved in ethanol at concentrations 1000-fold more than the concentration desired for cell treatments. Cells (106) are suspended in 1 ml of complete medium in microcentrifuge tubes; 1 μl of ethanol or of the ethanolic solution of Cyclosporin A is added, and the cells are incubated at 37°C for 1 hr. Cells are washed twice with 1 ml of PBS on ice and lysed in 50μl of hypotonic buffer containing 50 mM Tris (pH 7.5); 0.1 mM EGTA; 1 mM EDTA; 0.5 mM dithiothreitol; and 50 μg of phenylmethylsulfonyl fluoride, 50 μg of soybean trypsin inhibitor, 5 μg of leupeptin, and 5 μg of aprotinin per ml. Lysates are subjected to three cycles of freezing in liquid nitrogen followed by thawing at 30°C and then are centrifuged at 4°C for 10 min at 12,000×g.

    In VivoCyclosporin A is an immunosuppressive agent that is effective following either parenteral or oral administration in mice, rat and guinea pigs, is clinically used in organ transplantation to prevent rejection. 
    Animal modelRat 
    Formulation & DosageDissolved in 0.5% solution of tragacanth; 45 mg/kg; oral gavage
    References

    Science. 1984 Nov 2;226(4674):544-7; Cell. 1991 Aug 23;66(4):807-15; Biol Reprod. 2008 Jun;78(6):1102-10.  


    These protocols are for reference only. InvivoChem does not independently validate these methods.

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