Cyanide: Difference between revisions
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== Application in [[HRR]] == | == Application in [[HRR]] == | ||
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::::* [[alternative oxidase]] | ::::* [[alternative oxidase]] | ||
:::: ''' | :::: <span style="color:#8B008B"> '''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 ยตl. This now have been corrected to our previous recommendation of 1 M KCN stock solutions </span> | ||
::: '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | |||
::::* Titration volume: 2 ยตl of 1 M stock, using a 10 ยตl syringe (2 ml O2k-chamber). | |||
::::* Final concentration: 1 mM. | |||
=== Preparation of 1 M KCN stock === | |||
:::: '''Kcn'''(KCN): Fluka 60178, 100 g, store at R.T.; FW = 65.12. | |||
:::: ''' | :::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; KCN is volatile and very toxic! </span> | ||
:::: | :::: ''' Preparation of 1 M KCN stock solution ''' for '''2-mL O2k-chamber''' | ||
::::# Weigh 13 mg of KCN. | |||
::::''' | ::::# Dissolve in 0.2 ml H<sub>2</sub>O. | ||
::::# The pH of the solution may be very alkaline - adjust to physiological pH with HCl. | |||
ย ย ย ย ย :::ยป '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | |||
::::* Titration volume ('''2-mL O2k-chamber'''): 100 ยตL using a 100 ยตL Hamilton syringe. | |||
::::* Final concentration: 1 mM. | ::::* Final concentration: 1 mM. | ||
:::: '''Preparation of 250 mM KCN stock solution''' (in 45 mM Trizma base) for '''0.5-mL O2k-chamber''': | :::: '''Preparation of 250 mM KCN stock solution''' (in 45 mM Trizma base) for '''0.5-mL O2k-chamber''': |
Revision as of 12:40, 22 January 2021
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
- 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 ยตl. This now have been corrected to our previous recommendation of 1 M KCN stock solutions
- O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume: 2 ยตl of 1 M stock, using a 10 ยตl syringe (2 ml O2k-chamber).
- Final concentration: 1 mM.
- O2k manual titrations MiPNet09.12 O2k-Titrations
Preparation of 1 M 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.
:::ยป O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 100 ยตL using a 100 ยตL Hamilton syringe.
- Final concentration: 1 mM.
- Preparation of 250 mM KCN stock solution (in 45 mM Trizma base) for 0.5-mL O2k-chamber:
- Weigh 16,28 mg of KCN and dissolve in 1 mL of 45 mM Trizma base.
- Divide into 200 ยตL aliquots (use dark glass vials).
- Store frozen at -20 ยฐC.
- Preparation of 250 mM KCN stock solution (in 45 mM Trizma base) for 0.5-mL O2k-chamber:
- ยป O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (0.5-mL O2k-chamber): 2 ยตL using a 10 ยตL Hamilton syringe.
- Final concentration: 1 mM.
- ยป O2k manual titrations MiPNet09.12 O2k-Titrations
MitoPedia topics:
Inhibitor