Description
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 ยฐC: 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 ยฐC.
- 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 ยตl (or 0.5 ยตl) steps using a 10 ยตl syringe (2 ml O2k-Chamber).
- Final concentration: 0.5 ยตM (or 0.25 ยตM) 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]
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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
- Specific
- Respiratory states and control ratios
- ยป Biochemical coupling efficiency
- ยป Coupling-control state
- ยป Electron-transfer-pathway state
- ยป Electron-transfer pathway
- 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
- Respiratory states and control ratios
- Gnaiger E et al โ MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. https://doi.org/10.26124/bec:2020-0001.v1
- Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2. https://doi.org/10.26124/bec:2020-0002
- General (alphabetical order)
- Other keyword lists
- โ 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. ยปBioblast linkยซ
MitoPedia topics:
Uncoupler