Cyclosporin A: Difference between revisions
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:::: '''Preparation of 400 ยตM stock solution''' (dissolved in | :::: '''Preparation of 400 ยตM stock solution''' (dissolved in ethanol abs.) | ||
::::# Weigh 1 mg of CsA and dissolve in 2.11 ml of ethanol | ::::# Weigh 1 mg of CsA and dissolve in 2.11 ml of ethanol abs.. | ||
::::# Divide in aliquots. ย | ::::# Divide in aliquots. ย | ||
::::# Store frozen at -20 ยฐC. | ::::# Store frozen at -20 ยฐC. |
Revision as of 11:56, 22 June 2021
Description
Cyclosporin A (CsA) is a cyclic undecapeptide from an extract of soil fungi that binds the cyclophilin D and thus preventing the formation of the mitochondrial permeability transition pore. The interaction of CsA with the cyclophilin D is phosphate mediated but the full mechanism of interaction is not well understood. For example, the deficiency of cyclophilin D in KO models does not prevent mitochondria from permeability transition and from CsA inhibition. Moreover, it is also a is a calcineurin inhibitor and potent immunosuppressive agent used largely as a means of prophylaxis against cellular rejection after solid organ transplantation.
Abbreviation: CsA
Application in HRR
- CsA:Cyclosporin A (C62H111N11O12); Sigma 30024, 25 mg, store commercial stock at 2-8ยฐC; M = 1202.61 gยทmol-1 (โฅ 95%).
- Preparation of 400 ยตM stock solution (dissolved in ethanol abs.)
- Weigh 1 mg of CsA and dissolve in 2.11 ml of ethanol abs..
- Divide in aliquots.
- Store frozen at -20 ยฐC.
- ยป O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 5 ยตL using a 10 ยตL syringe (2 mL O2k-chamber).
- Final concentration: 1 ยตM.
Even if 1 ยตM is recommended as the working concentration in most of the publications it is highly relevant to test the optimal concentration for your research model.
Inhibition of PTP by CsA
- Gainutdinov T, Molkentin JD, Siemen D, Ziemer M, Debska-Vielhaber G, Vielhaber S, Gizatullina Z, Orynbayeva Z, Gellerich FN (2015) Knockout of cyclophilin D in Ppif-/- mice increases stability of brain mitochondria against Ca2+ stress. Arch Biochm Biophys 579:40โ46.
- Briston T, Roberts M, Lewis S, Powney B, Staddon JM, Szabadkai G, Duchen MR (2017) Mitochondrial permeability transition pore: sensitivity to opening and mechanistic dependence on substrate availability. Scientific reports 7: 10492.
MitoPedia topics: Inhibitor