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
Inhibitor titrations, CIV-assay, alternative oxidase and ROX
- Cyanide and determination of residual oxygen consumption (ROX).
- Complex IV assay
- alternative oxidase
Preparation of 20 mM KCN stock (in 50mM Trizma base)
- Kcn(KCN): Fluka 60178, 100 g, store at R.T.; FW = 65.12., Trizma base: Sigma T6066, 100 g, store at R.T.; FW = 121.14
- Caution: KCN is photosensitive and hygroscopic; HCN is volatile and very toxic! It is recommended to prepared fresh although it can be safely stored at -20Β°C. To avoid KCN evaporation, it is recommended to use alkaline buffers (such as 50 mM Trizma base, pH 9).
- Preparation of 50 mM Trizma base:
- Weigh 605.7 mg of Trizma base
- Dissolve it in 100 mL of H20.
- Preparation of 50 mM Trizma base:
- Preparation of 20 mM KCN:
- Weigh 5.2 mg of KCN and dissolve in 4 ml of 50 mM Trizma base.
- Divide into 500 Β΅l aliquots (use dark glass vials).
- Store frozen at -20 Β°C.
- Preparation of 20 mM KCN:
- O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 100 Β΅l of 20 mM stock (2 ml O2k-chamber).
- Final concentration: 1 mM.
- O2k manual titrations MiPNet09.12 O2k-Titrations