Rotenone: Difference between revisions
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::::* Titration volume: 1 ยตl using a 10 ยตl syringe (2 ml O2k-chamber). | ::::* Titration volume: 1 ยตl using a 10 ยตl syringe (2 ml O2k-chamber). | ||
::::* Final concentration: 0.5 ยตM. | ::::* Final concentration: 0.5 ยตM. | ||
'''Microalgae applications''' | '''Microalgae applications''' |
Revision as of 16:48, 10 April 2019
Description
Rotenone is an inhibitor of Complex I (CI) and thus inhibits NADH oxidation. It inhibits the transfer of electrons from iron-sulfur centers in CI to ubiquinone via binding to the ubiquinone binding site of CI. See also Complex II-linked substrate state, S(Rot).
Abbreviation: Rot
Reference: Gnaiger 2014 MitoPathways, MiPNet09.12, MiPNet03.02
Application in HRR
- Rot: Rotenone (C23H22O6); Sigma R 8875, 1 g, store commercial stock at RT; FW = 394.42 (โฅ 95%).
- Caution: Very toxic and light sensitive (store solution in a dark glass vial)..
- Preparation of 1 mM stock solution (dissolved in 99.9% ethanol)
- Weigh 0.39 mg of rotenone and dissolve in 1 ml of ethanol absolute (99.9%).
- Divide into 100 ยตl aliquots (use dark glass vials).
- Store frozen at -20 ยฐC.
- O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 1 ยตl using a 10 ยตl syringe (2 ml O2k-chamber).
- Final concentration: 0.5 ยตM.
Microalgae applications
- For the specific case of microalgal cells it is recommended to prepare higher concentration:
- Preparation of 5.5 mM stock solution (dissolved in 99.9% ethanol)
- Weigh 4.4 mg of rotenone and dissolve in 2 ml of ethanol absolute (99.9%).
- Divide into 200 ยตl aliquots (use dark glass vials).
- Store frozen at -20 ยฐC.
Inhibition of CI by rotenone
- Fato R, Bergamini C, Bortolus M, Maniero AL, Leoni S, Ohnishi T, Lenaz G (2009) Differential effects of mitochondrial Complex I inhibitors on production of reactive oxygen species. Biochim Biophys Acta 1787:384โ92. >>PMC2724837
MitoPedia topics: Inhibitor