Difference between revisions of "Cyanide"
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=== Preparation of 20 mM KCN stock (in | === Preparation of 20 mM KCN stock (in 45mM 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 | :::: '''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 | ||
:::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; HCN is volatile and very toxic! It is recommended to prepare fresh KCN solution although it can be safely stored at -20°C. To avoid KCN evaporation, it is recommended to use alkaline buffers (such as | :::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; HCN is volatile and very toxic! It is recommended to prepare fresh KCN solution although it can be safely stored at -20°C. To avoid KCN evaporation, it is recommended to use alkaline buffers (such as 45 mM Trizma base, pH >10).</span> | ||
:::: '''Preparation of | :::: '''Preparation of 45 mM Trizma base:''' | ||
::::# Weigh | ::::# Weigh 545 mg of Trizma base | ||
::::# Dissolve it in 100 mL of H<sub>2</sub>0. | ::::# Dissolve it in 100 mL of H<sub>2</sub>0. | ||
:::: '''Preparation of 20 mM KCN:''' | :::: '''Preparation of 20 mM KCN:''' | ||
::::# Weigh 5.2 mg of KCN and dissolve in 4 ml of | ::::# Weigh 5.2 mg of KCN and dissolve in 4 ml of 45 mM Trizma base. | ||
::::# Divide into 500 µl aliquots (use dark glass vials). | ::::# Divide into 500 µl aliquots (use dark glass vials). | ||
::::# Store frozen at -20 °C. | ::::# Store frozen at -20 °C. |
Revision as of 15:53, 19 April 2019
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 45mM 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 prepare fresh KCN solution although it can be safely stored at -20°C. To avoid KCN evaporation, it is recommended to use alkaline buffers (such as 45 mM Trizma base, pH >10).
- Preparation of 45 mM Trizma base:
- Weigh 545 mg of Trizma base
- Dissolve it in 100 mL of H20.
- Preparation of 45 mM Trizma base:
- Preparation of 20 mM KCN:
- Weigh 5.2 mg of KCN and dissolve in 4 ml of 45 mM Trizma base.
- Divide into 500 µl aliquots (use dark glass vials).
- Store frozen at -20 °C.
- Preparation of 20 mM KCN:
- O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 100 µl of 20 mM stock (2 ml O2k-chamber).
- Final concentration: 1 mM.
- O2k manual titrations MiPNet09.12 O2k-Titrations