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Difference between revisions of "Cyanide"

From Bioblast
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::::* [[alternative oxidase]]
::::* [[alternative oxidase]]


::: '''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.




=== Preparation of 1 M KCN stock ===
=== Preparation of 20 mM KCN stock (in 50mM 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
 
:::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; HCN is volatile and very toxic! Has to be prepared in 50 mM Trizma base to keep it in a high pH range and avoid any evaporation of the KCN </span>
 
:::: '''Preparation of 50 mM Trizma base:'''
::::# Weigh 605.7 mg of Trizma base
::::# Dissolve it in 100 mL of H<sub>2</sub>0.


:::: '''Kcn'''(KCN): Fluka 60178, 100 g, store at R.T.; FW = 65.12.
:::: '''Preparation of 20 mM KCN:'''  
::::# Weigh 5.2 mg of KCN and dissolve in 4 ml of 50 mM Trizma base.
::::# Divide into 500 µl aliquots (use dark glass vials).
::::# Store frozen at -20 °C.


:::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; HCN is volatile and very toxic! Prepare the solution fresh everyday!</span>


::::# Weigh 13 mg of KCN.  
::: '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
::::# Dissolve in 0.2 ml H<sub>2</sub>O.
::::* Titration volume: 100 µl of 20 mM stock (2 ml O2k-chamber).
::::# The pH of the solution may be very alkaline - adjust to physiological pH with HCl.
::::* Final concentration: 1 mM.

Revision as of 12:44, 12 April 2019


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



MitoPedia topics: Inhibitor 

Application in HRR

Inhibitor titrations, CIV-assay, alternative oxidase and ROX


Preparation of 20 mM KCN stock (in 50mM 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! Has to be prepared in 50 mM Trizma base to keep it in a high pH range and avoid any evaporation of the KCN
Preparation of 50 mM Trizma base:
  1. Weigh 605.7 mg of Trizma base
  2. Dissolve it in 100 mL of H20.
Preparation of 20 mM KCN:
  1. Weigh 5.2 mg of KCN and dissolve in 4 ml of 50 mM Trizma base.
  2. Divide into 500 µl aliquots (use dark glass vials).
  3. Store frozen at -20 °C.


O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume: 100 µl of 20 mM stock (2 ml O2k-chamber).
  • Final concentration: 1 mM.