Rotenone: Difference between revisions
From Bioblast
No edit summary |
No edit summary |
||
Line 13: | Line 13: | ||
== Application in [[HRR]] == | == Application in [[HRR]] == | ||
'''Rot: Rotenone''' (C<sub>23</sub>H<sub>22</sub>O<sub>6</sub>); Sigma R 8875, 1 g, store commercial stock at RT; FW = 394.42 (โฅ 95%). | :::: '''Rot: Rotenone''' (C<sub>23</sub>H<sub>22</sub>O<sub>6</sub>); Sigma R 8875, 1 g, store commercial stock at RT; FW = 394.42 (โฅ 95%). | ||
<span style="color:#8B008B"> '''Caution:''' Very toxic and light sensitive (store solution in a dark glass vial). Use fresh aliquots for every experimental day!</span>. | :::: <span style="color:#8B008B"> '''Caution:''' Very toxic and light sensitive (store solution in a dark glass vial). Use fresh aliquots for every experimental day!</span>. | ||
'''Preparation of 1 mM stock solution''' (dissolved in 99.9% ethanol) | ::: '''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). ย | ||
:: | ::::# Store frozen at -20 ยฐC. | ||
'''O2k manual titrations'''ย [[MiPNet09.12 O2k-Titrations]] | ::: '''O2k manual titrations'''ย [[MiPNet09.12 O2k-Titrations]] | ||
::* 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. | ||
== Inhibition of CI by rotenone == | == 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: | ::::* 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. [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2724837/ >>PMC2724837] |
Revision as of 18:06, 17 July 2016
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
MitoPedia topics: Inhibitor
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). Use fresh aliquots for every experimental day!.
- 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.
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