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 KCN stock
- KCN: Cyanide (Potassium cyanide; KCN), Sigma-Aldrich: 60178, store at RT, CAS: 151-50-8, M = 65.12 gΒ·mol-1
- Hazard statements: H290, H300, H330, H310, H372, H400, H410; may be corrosive to metals, fatal if swallowed, fatal if inhaled, fatal in contact with skin, causes damage to organs through prolonged or repeated exposure (thyroid), very toxic to aquatic life, very toxic to aquatic life with long-lasting effects
- KCN: Cyanide (Potassium cyanide; KCN), Sigma-Aldrich: 60178, store at RT, CAS: 151-50-8, M = 65.12 gΒ·mol-1
- Caution: Very toxic! Protect from light, store in a cool, dry, well-ventilated location. Separate from water, acids, and carbon dioxide. For details refer to PubChem
- Caution: In April 2019 a change in the procedure for preparing the KCN was done that recommended to prepare 20 mM KCN stock and titrate 100 Β΅L. 2021-01-22: after discussions on KCN toxicity we recommend to prepare of 0.5 M KCN stock solutions.
- Preparation of 0.5 M KCN stock solution (dissolved in H2O):
- Weigh 32.6 mg of KCN.
- Dissolve in 1 ml H2O.
- The pH of the solution is basic. Since the titration amount is low and HCN may evaporate during pH adjustment with HCl, the pH is not adjusted.
- Divide into 0.2 mL portions
- Store at -20 Β°C.
- Β» O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 4 Β΅L using a 10 Β΅L Hamilton syringe.
- Titration volume (0.5-mL O2k-chamber): 1 Β΅L using a 10 Β΅L Hamilton syringe.
- Final concentration: 1 mM.
- Β» O2k manual titrations MiPNet09.12 O2k-Titrations
MitoPedia topics: Inhibitor