Difference between revisions of "Cyanide"

Revision as of 12:07, 22 January 2021

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

Cyanide and determination of residual oxygen consumption (ROX).
Complex IV assay
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

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! Prepare the solution fresh everyday!

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.

@@ Line 4: / Line 4: @@
 |info=[[MiPNet09.12]]
 }}
+{{MitoPedia O2k and high-resolution respirometry|type=Inhibitor ETS
+}}
+{{MitoPedia topics
+|mitopedia topic=Inhibitor
+}}
 __TOC__
 == Application in [[HRR]] ==
@@ Line 13: / Line 17: @@
 ::::* [[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
+::: '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
+::::* Titration volume: 2 µl of 1 M stock, using a 10 µl syringe (2 ml O2k-chamber).
-:::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; KCN 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>
+::::* Final concentration: 1 mM.
-:::: '''Preparation of 45 mM Trizma base:'''
-::::# Weigh 545 mg of Trizma base
-::::# Dissolve it in 100 mL of H<sub>2</sub>0.
-::::''' Preparation of 20 mM KCN stock solution ''' (in 45 mM Trizma base) for '''2-mL O2k-chamber''':
+=== Preparation of 1 M KCN stock ===
-::::# 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.
-:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]
+:::: '''Kcn'''(KCN): Fluka 60178, 100 g, store at R.T.; FW = 65.12.
-::::* Titration volume ('''2-mL O2k-chamber'''): 100 µL using a 100 µL Hamilton syringe.
-::::* Final concentration: 1 mM.
+:::: <span style="color:#8B008B"> '''Caution:''' KCN is photosensitive and hygroscopic; HCN is volatile and very toxic! Prepare the solution fresh everyday!</span>
-:::: '''Preparation of 250 mM KCN stock solution''' (in 45 mM Trizma base) for '''0.5-mL O2k-chamber''':
+::::# Weigh 13 mg of KCN.
-::::# Weigh 16,28 mg of KCN and dissolve in 1 mL of 45 mM Trizma base.
+::::# Dissolve in 0.2 ml H<sub>2</sub>O.
-::::# Divide into 200 µL aliquots (use dark glass vials).
+::::# The pH of the solution may be very alkaline - adjust to physiological pH with HCl.
-::::# Store frozen at -20 °C.
-:::» '''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.
-{{MitoPedia O2k and high-resolution respirometry|type=Inhibitor ETS
-}}
-{{MitoPedia topics
-|mitopedia topic=Inhibitor
-}}