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FCCP

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Revision as of 05:12, 22 April 2014 by Gnaiger Erich (talk | contribs)


high-resolution terminology - matching measurements at high-resolution


FCCP

Description

FCCP (Carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone) is a protonophore or uncoupler: FCCP added at uncoupler concentration Uc; c is the optimum uncoupler concentration in titrations to obtain maximum mitochondrial respiration in the noncoupled state of ETS capacity.

Abbreviation: Uc

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



MitoPedia topics: Uncoupler 

CCCP versus FCCP

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

  • FCCP should be used instead of CCCP at low experimental temperature, or in laboratories at low room temperature that may prevail during expeditions, to avoid blocking the needle of the Hamilton titration syringe.


Application in HRR

F: FCCP (Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, C10H5F3N4O); Sigma C 2920, 10 mg, store at 4 °C: FW = 254.17)


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.

Oxygraph-2k 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]

  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: 327-352. »Bioblast Access