Cyanide: Difference between revisions
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|info=[[MiPNet09.12]] | |info=[[MiPNet09.12]] | ||
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== Application in [[HRR]] == | == Application in [[HRR]] == | ||
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::::* [[alternative oxidase]] | ::::* [[alternative oxidase]] | ||
:::: '''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 | |||
:::: <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> | :::: <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 20 mM KCN stock (in 45 mM Trizma base)''' | |||
:::: '''Preparation of 45 mM Trizma base:''' | :::: '''Preparation of 45 mM Trizma base:''' | ||
::::# Weigh 545 mg of Trizma base | ::::# Weigh 545 mg of Trizma base | ||
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:::: '''Preparation of 20 mM KCN:''' | :::: '''Preparation of 20 mM KCN:''' | ||
::::# Weigh 5.2 mg of KCN and dissolve in 4 mL of 45 mM Trizma base. | ::::# Weigh 5.2 mg of KCN and dissolve in 4 mL of 45 mM Trizma base. | ||
::::# Divide into 500 | ::::# Divide into 500 µL aliquots (use dark glass vials). | ||
::::# Store frozen at -20 °C. | ::::# Store frozen at -20 °C. | ||
::: '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | :::» '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
::::* Titration volume: 100 µL of 20 mM stock (2 mL O2k-chamber). | ::::* Titration volume: 100 µL of 20 mM stock (2 mL O2k-chamber). | ||
::::* Final concentration: 1 mM. | ::::* Final concentration: 1 mM. | ||
{{MitoPedia O2k and high-resolution respirometry|type=Inhibitor ETS | |||
}} | |||
{{MitoPedia topics | |||
|mitopedia topic=Inhibitor | |||
}} | |||
Revision as of 10:17, 11 September 2020
- high-resolution terminology - matching measurements at high-resolution
Cyanide
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
Application in HRR
Inhibitor titrations, CIV-assay, alternative oxidase and ROX
- Cyanide and determination of residual oxygen consumption (ROX).
- Complex IV assay
- alternative oxidase
- 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 20 mM KCN stock (in 45 mM Trizma base)
- Preparation of 45 mM Trizma base:
- Weigh 545 mg of Trizma base
- Dissolve it in 100 mL of H20.
- 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
MitoPedia topics:
Inhibitor
