Difference between revisions of "Cyanide"
From Bioblast
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::::* Final concentration: 1 mM. | ::::* Final concentration: 1 mM. | ||
=== Preparation of | === Preparation of KCN stock === | ||
:::: '''Kcn'''(KCN): Fluka 60178, 100 g, store at R.T.; FW = 65.12. | :::: '''Kcn'''(KCN): Fluka 60178, 100 g, store at R.T.; FW = 65.12. | ||
Line 32: | Line 32: | ||
::::# Weigh 13 mg of KCN. | ::::# Weigh 13 mg of KCN. | ||
::::# Dissolve in 0.2 ml H<sub>2</sub>O. | ::::# 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. | ::::# The pH of the solution may be very alkaline - adjust to physiological pH with HCl. |
Revision as of 12:44, 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
Last update 2021-01-22
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 µl. 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 µl of 1 M stock, using a 10 µl syringe (2 ml O2k-chamber).
- Final concentration: 1 mM.
- O2k manual titrations MiPNet09.12 O2k-Titrations
Preparation of 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.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 100 µL using a 100 µL 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 µL aliquots (use dark glass vials).
- Store frozen at -20 °C.
- Preparation of 250 mM KCN stock solution (in 45 mM Trizma base) for 0.5-mL O2k-chamber:
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (0.5-mL O2k-chamber): 2 µL using a 10 µL Hamilton syringe.
- Final concentration: 1 mM.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
MitoPedia topics:
Inhibitor