Carbonyl cyanide m-chlorophenyl hydrazone: Difference between revisions
From Bioblast
No edit summary |
No edit summary |
||
| Line 17: | Line 17: | ||
=== MiPNet News CCCP 2013-07-11 === | === MiPNet News CCCP 2013-07-11 === | ||
: '''We replace [[FCCP]] by CCCP in the OROBOROS Mitochondrial Research Laboratory'''. Up to now, we used and recommended FCCP as an uncoupler in high- | : '''We replace [[FCCP]] by CCCP in the OROBOROS Mitochondrial Research Laboratory'''. Up to now, we used and recommended FCCP as an uncoupler in high-resolution respirometry. Recently we compared the uncoupling effect of CCCP and FCCP in our on-going experiments with homogenate preparations of mouse brain and heart. In parallel experiments identical results were obtained on ETS capacity and mt-membrane potential. Compared to FCCP, a much higher [[DNP]] concentration is required for maximum stimulation of oxygen flux ([[Steinlechner-Maran 1996 Am J Physiol Cell Physiol]]). In contrast, the optimum CCCP concentration for maximum oxygen flux was only slightly higher than the optimum FCCP concentration. '''CCCP is significantly less expensive than FCCP'''. - [[Eigentler A]], [[Gnaiger E]] | ||
Revision as of 08:42, 8 August 2013
- high-resolution terminology - matching measurements at high-resolution
Carbonyl cyanide m-chlorophenyl hydrazone
Description
Carbonyl cyanide m-chloro phenyl hydrazone (C9H5ClN4; FW = 204.62) is a protonophore (H+ ionophore) and is used as a potent chemical uncoupler of oxidative phosphorylation.
Abbreviation: CCCP
Reference: Lou 2007 Biochem J, Skulachev 1998 Biochim Biophys Acta
MitoPedia methods:
Respirometry
MitoPedia topics:
Uncoupler
Application in HRR
MiPNet News CCCP 2013-07-11
- We replace FCCP by CCCP in the OROBOROS Mitochondrial Research Laboratory. Up to now, we used and recommended FCCP as an uncoupler in high-resolution respirometry. Recently we compared the uncoupling effect of CCCP and FCCP in our on-going experiments with homogenate preparations of mouse brain and heart. In parallel experiments identical results were obtained on ETS capacity and mt-membrane potential. Compared to FCCP, a much higher DNP concentration is required for maximum stimulation of oxygen flux (Steinlechner-Maran 1996 Am J Physiol Cell Physiol). In contrast, the optimum CCCP concentration for maximum oxygen flux was only slightly higher than the optimum FCCP concentration. CCCP is significantly less expensive than FCCP. - Eigentler A, Gnaiger E
Preparation
- C: CCCP (Carbonyl cyanide 3-chlorophenylhydrazone, C9H5ClN4O); Sigma C 2759, 100 mg, store at -20 °C: FW = 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.
