Carbonyl cyanide m-chlorophenyl hydrazone: Difference between revisions

Revision as of 09:40, 11 July 2013

high-resolution terminology - matching measurements at high-resolution

Carbonyl cyanide m-chlorophenyl hydrazone

Description

Carbonyl cyanide m-chloro phenyl hydrazone (C₉H₅ClN₄; F_W = 204.62) is a protonophore (H⁺ ionophore) and is used as a chemical uncoupler of oxidative phosphorylation.

Abbreviation: CCCP

MitoPedia methods: Respirometry

MitoPedia topics: Uncoupler

Labels:

Regulation: Respiration; OXPHOS; ETS Capacity"Respiration; OXPHOS; ETS Capacity" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.

HRR: Chemicals; Media"Chemicals; Media" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property.

Application in HRR

MiPNet News (2013-07-11)

Using CCCP as an uncoupler in our on-going experiments with homogenate preparations of mouse brain and heart, we obtained results on maximum ETS capacity and mt-membrane potential which were identical to parallel experiments using FCCP. Therefore, we replace in our Mitochondrial Research Laboratory FCCP by CCCP. Importantly, CCCP is significantly less expensive than the most frequently used uncoupler FCCP. The optimum CCCP concentration for maximum oxygen flux is only slightly higher than the optimum FCCP concentration. - Eigentler A, Gnaiger E

Preparation

C: CCCP (Carbonyl cyanide 3-chlorophenylhydrazone, C₉H₅ClN₄O); Sigma C 2759, 100 mg, store at -20 °C: F_W = 204.62

Preparation of 1 mM stock solution (dissolved in DMSO):

1) Weigh 1.02 mg of CCCP into a glass vial.

2) Dissolve in 5 ml DMSO.

3) Divide into 0.2 ml portions OR store in 5 ml glass vial and use subsamples during experiments to avoid contamination of the stock solution.

4) Store in glass vials at -20 °C.

5) Titrations: 10 µl syringe, 1 µl steps, corresponding to 0.5 µM steps of the concentration in the 2 ml O2k-chamber.

Caution: Toxic!

Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=CCCP
-|description='''Carbonyl cyanide m-chloro phenyl hydrazone''' (C<sub>9</sub>H<sub>5</sub>ClN<sub>4</sub>; FW = 204.62) is a protonophore (H<sup>+</sup> ionophore) and chemical uncoupler of [[Oxidative phosphorylation|oxidative phosphorylation]]. CCCP causes uncoupling ([[uncoupler]]) of the proton gradient that is established during the normal activity of electron carriers in the [[Electron transport system|electron transport system]].
+|description='''Carbonyl cyanide m-chloro phenyl hydrazone''' (C<sub>9</sub>H<sub>5</sub>ClN<sub>4</sub>; ''F''<sub>W</sub> = 204.62) is a protonophore (H<sup>+</sup> ionophore) and is used as a chemical [[uncoupler]] of [[oxidative phosphorylation]].
+|type=Uncoupler
+}}
+{{MitoPedia methods
+|mitopedia method=Respirometry
 |type=Uncoupler
 }}
@@ Line 18: / Line 22: @@
 == Application in [[HRR]] ==
-'''C: CCCP''' (Carbonyl cyanide 3-chlorophenylhydrazone, C<sub>9</sub>H<sub>5</sub>ClN<sub>4</sub>O);  Sigma C 2759, 100 mg, store at -20 °C: FW = 204.62
+=== MiPNet News (2013-07-11) ===
+: Using CCCP as an uncoupler in our on-going experiments with homogenate preparations of mouse brain and heart, we obtained results on maximum ETS capacity and mt-membrane potential which were identical to parallel experiments using [[FCCP]]. Therefore, '''we replace in our Mitochondrial Research Laboratory FCCP by CCCP'''. Importantly, CCCP is significantly less expensive than the most frequently used uncoupler FCCP. The optimum CCCP concentration for maximum oxygen flux is only slightly higher than the optimum FCCP concentration. - [[Eigentler A]], [[Gnaiger E]]
-<span style="color:#8B008B"> '''Caution:''' Toxic!</span>
-<span  style="color:#8B008B"> '''Caution:''' Chemicals stored in the fridge  or freezer should be allowed to reach room temperature before  opening.</span>
+=== Preparation ===
+: '''C: CCCP''' (Carbonyl cyanide 3-chlorophenylhydrazone, C<sub>9</sub>H<sub>5</sub>ClN<sub>4</sub>O);  Sigma C 2759, 100 mg, store at -20 °C: ''F''<sub>W</sub> = 204.62
+: '''Preparation of 1 mM stock solution''' (dissolved in DMSO):
-'''Preparation of 1 mM stock solution''' (dissolved in DMSO):
+::1) Weigh 1.02 mg of CCCP into a glass vial.
+::2) Dissolve in 5 ml DMSO.
+::3) Divide into 0.2 ml portions OR store in 5 ml glass vial and use subsamples during experiments to avoid contamination of the stock solution.
+::4) Store in glass vials at -20 °C.
+::5) Titrations: 10 µl syringe, 1 µl steps, corresponding to 0.5 µM steps of the concentration in the 2 ml O2k-chamber.
-::1) weigh 1.02 mg of CCCP into a glass vial
-::2) dissolve in 5 ml DMSO
-::3) divide into 0.2 ml portions OR store in 5 ml glass vial and use subsamples during experiments to avoid contamination of the stock solution
-::3) store in glass vials at -20 °C
-::* Titration volume: 1 µl steps using a 10 µl syringe
+<span style="color:#8B008B"> '''Caution:''' Toxic!</span>
-::* Final conc. in 2 ml O2k-chamber: 0.5 µM steps
+<span  style="color:#8B008B"> '''Caution:''' Chemicals stored in the fridge  or freezer should be allowed to reach room temperature before  opening.</span>