From Bioblast
Description
Cyanide (usually added as KCN) is a competitive inhibitor of cytochcrome c oxidase (CIV). Inhibition is reversed by pyruvate and high oxygen levels.
Abbreviation: KCN
Reference: MiPNet09.12
MitoPedia topics: Inhibitor
Application in HRR
Last update 2021-01-22
Inhibitor titrations, CIV-assay, alternative oxidase and ROX
- Cyanide and determination of residual oxygen consumption (ROX).
- Complex IV assay
- alternative oxidase
- Caution: Due to a confusion, a change in the procedure for preparing the KCN was done in April 2019 that recommended to prepare 20 mM KCN stock and titrate 100 Β΅l. This now have been corrected to our previous recommendation of 1 M KCN stock solutions
Preparation of KCN stock
- Kcn(KCN): Fluka 60178, 100 g, store at R.T.; FW = 65.12.
- Caution: KCN is photosensitive and hygroscopic; KCN is volatile and very toxic!
- Preparation of 1 M KCN stock solution for 2-mL O2k-chamber
- Weigh 13 mg of KCN.
- Dissolve in 0.2 ml H2O.
- The pH of the solution may be very alkaline - adjust to physiological pH with HCl.
- Β» O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 100 Β΅L using a 100 Β΅L Hamilton syringe.
- Final concentration: 1 mM.
MitoPedia topics: Inhibitor