Difference between revisions of "Cyanide"
From Bioblast
(changed concentration of stock solution from 1 M to 0.5 M. Now it can be used for 2ml- as well as for 0.5ml-chambers; refined instructions (after clarification with Erich)) |
|||
Line 25: | Line 25: | ||
:::: '''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. | ||
:::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; | :::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; HCN is volatile and very toxic! </span> | ||
:::: ''' Preparation of | |||
:::: ''' Preparation of 0.5 M KCN stock solution''' (dissolved in H<sub>2</sub>O): | |||
::::# Weigh 32.6 mg of KCN. | |||
::::# Dissolve in 1 ml H<sub>2</sub>O. | |||
::::# 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 °C. | |||
:::» '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | :::» '''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ||
::::* Titration volume ('''2-mL O2k-chamber'''): | ::::* Titration volume ('''2-mL O2k-chamber'''): 4 µL using a 10 µL Hamilton syringe. | ||
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL using a 10 µL Hamilton syringe. | |||
::::* Final concentration: 1 mM. | ::::* Final concentration: 1 mM. | ||
Revision as of 13:12, 29 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 has been corrected to our previous recommendation of 1 M KCN stock solutions
Preparation of KCN stock
- Kcn(KCN): Fluka 60178, 100 g, store at R.T.; FW = 65.12.
- Caution: KCN is photosensitive and hygroscopic; HCN is volatile and very toxic!
- 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 °C.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
- Titration volume (2-mL O2k-chamber): 4 µL using a 10 µL Hamilton syringe.
- Titration volume (0.5-mL O2k-chamber): 1 µL using a 10 µL Hamilton syringe.
- Final concentration: 1 mM.
- » O2k manual titrations MiPNet09.12 O2k-Titrations
MitoPedia topics: Inhibitor