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Bioblasts - Richard Altmann and MiPArt by Odra Noel
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FCCP (Carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone, C10H5F3N4O) is a protonophore or uncoupler: added at uncoupler concentration Uc; c is the optimum uncoupler concentration in titrations to obtain maximum mitochondrial respiration in the noncoupled state of ET capacity.

Abbreviation: FCCP

Reference: Steinlechner-Maran 1996 Am J Physiol Cell Physiol, Gnaiger_2008_POS

CCCP versus FCCP

Bioblast alert 2013(02): We replace FCCP by the less expensive and equally effective CCCP in our experiments with high-resultion respirometry (HRR).

Application in HRR

FCCP (Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, C10H5F3N4O); Sigma C 2920, 10 mg, store at 4 °C: M = 254.17 g·mol-1)
Caution: Toxic!

Preparation of 1 mM stock solution (dissolved in 99.9% ethanol):
1. Weigh 1.27 mg of FCCP into a glass vial.
2. Dissolve in 5 ml ethanol.
3. Divide into 0.2 ml portions, or store in 5 ml glass vial and use subsamples during experiments to avoid contamination of the storage solution.
4. Store in glass vials at -20 °C.
Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.
O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume: 1 µl (or 0.5 µl) steps using a 10 µl syringe (2 ml O2k-Chamber).
  • Final concentration: 0.5 µM (or 0.25 µM) steps.
Frequently, a more diluted FCCP stock is necessary for reducing the concentration steps in manual titrations. In this case prepare more diluted stocksolution (ditution 2-10x) from your storage solution. The highest accuracy of uncoupler titrations is achieved by titrations with the TIP2k at high concentrations of the stock solution. [1]


Click to expand or collaps

  1. Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial dysfunction in drug-induced toxicity (Dykens JA, Will Y, eds) John Wiley:327-52. »Bioblast link«

MitoPedia topics: Uncoupler