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Carbonyl cyanide m-chlorophenyl hydrazone

From Bioblast


high-resolution terminology - matching measurements at high-resolution


Carbonyl cyanide m-chlorophenyl hydrazone

Description

Carbonyl cyanide m-chlorophenyl hydrazone, CCCP (U; C9H5ClN4; FW = 204.62) is a protonophore (H+ ionophore) and is used as a potent chemical uncoupler of oxidative phosphorylation. Like all uncouplers, CCCP concentrations must be titrated carefully to evaluated the optimum concentration for maximum stimulation of mitochondrial respiration, particularly to avoid inhibition of respiration at higher CCCP concentrations.

Abbreviation: CCCP

Reference: Lou 2007 Biochem J, Skulachev 1998 Biochim Biophys Acta

Application in HRR

C: CCCP (Carbonyl cyanide 3-chlorophenylhydrazone, C9H5ClN4O); Sigma C 2759, 100 mg, store at -20 ยฐC: FW = 204.62. Recommended due to better handling in room temperature, we use for preparation ethanol in OROBOROS Lab.
Caution: Toxic!
Preparation of 1 mM stock solution (dissolved in Ethanol):
  1. Weigh 1.02 mg of CCCP into a glass vial.
  2. Dissolve in 5 mL ethanol (or DMSO - see note below).
  3. Store in glass vials at -20 ยฐC. Bring to room temperature before opening.
  • Storage options:
  1. Divide into 0.2 mL portions in glass vials.
  2. Store the whole solution in 5 mL glass vial and use subsamples during experiments to avoid contamination of the stock solution.


Solubility: CCCP also can be prepared in DMSO. Soluble in DMSO (5 mg/mL), ethanol (1 mg/mL), and methanol (10 mg/mL). Insoluble in water. DMSO has a freezing point of +18.55 ยฐC (65.4 ยฐF).
CCCP is soluble in ethanol up to 10 mM. Working with CCCP below room temperature is therefore possible. - Heidler J
O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume: 1 ยตL (or 0.5 ยตL) steps using a 10 ยตL syringe (2 mL O2k-chamber).
  • Final concentration: 0.5 (or 0.25 ยตM) ยตM steps
Caution: Bring to room temperature before filling the 10 ยตL syringe, which is important to avoid blocking the needle.


NOTE: 0.1 mM stock for mt-preparations with high uncoupler sensitivity; 1 mM stock for mt-preparations with low uncoupler sensitivity, livingcells invarious culture media (e.g.,RPMI, DMEM, EGC) and for TIP2k.

MiPNet News CCCP 2019-05-02

CCCP solution is less yellow coloured than FCCP. This may be important when working with fluorometric applications in parallel with high-resolution respirometry. - Cardoso LHD


MiPNet News CCCP 2014-01-15

CCCP is soluble in ethanol up to 10 mM. Working with CCCP below room temperature is therefore possible. - Heidler J


MiPNet News CCCP 2014-01-10

The temperature of the CCCP stock solution dissolved in DMSO should be increased to room temperature before filling the titration syringe. When the CCCP stock solution is kept on ice, the needle of the Hamilton titration syringe can be easily blocked, causing problems during titrations which may not be detected immediately. When performing experiments below room temperature, the needle of the Hamilton syringe is regularly blocked during injections into the O2k chamber. To avoid such problems, FCCP should be used instead of CCCP at low experimental temperature, or in laboratories at low room temperature that may prevail during expeditions. - Laner V, Gnaiger E


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 ET 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
  • Bioblast alert 2013(02)





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Specific
ยป Artefacts by single dose uncoupling
ยป ATP synthase
ยป CCCP
ยป Coupling-control protocol
ยป DNP
ยป Dyscoupled respiration
ยป FCCP
ยป Is respiration uncoupled - noncoupled - dyscoupled?
ยป Noncoupled respiration: Discussion
ยป Uncoupler
ยป Uncoupled respiration - see ยป Noncoupled respiration
ยป Uncoupling proteins
ยป Uncoupling protein 1
ยป Uncoupler titrations - Optimum uncoupler concentration
Respiratory states and control ratios
ยป Biochemical coupling efficiency
ยป Coupling-control state
ยป Electron-transfer-pathway state
ยป Electron-transfer pathway
E.jpg ET capacity
ยป E-L coupling efficiency
ยป Flux control efficiency
ยป Flux control ratio
ยป LEAK-control ratio
ยป LEAK respiration
ยป Noncoupled respiration
ยป OXPHOS
ยป OXPHOS capacity; ยป State 3
ยป OXPHOS-control ratio, P/E ratio
ยป Respiratory acceptor control ratio
ยป ROUTINE-control ratio
ยป ROUTINE respiration
ยป ROUTINE state
ยป State 3u
ยป State 4
ยป Uncoupling-control ratio UCR
General (alphabetical order)
ยป Adenine nucleotide translocase
ยป Adenylates
ยป Electron transfer pathway
ยป Mitochondrial preparations
ยป mt-membrane potential
ยป Oxygen flux
ยป Phosphorylation system
ยป Proton leak
ยป Proton slip
ยป TIP2k
Other keyword lists
ยป Template:Keywords: Force and membrane potential



MitoPedia topics: Uncoupler