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: MiPNet10.04; SteinlechnerMaran_1996_AJP
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).
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!
Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.
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 stock solution.
- 3) Store in glass vials (recommendation) at -20 °C.
Oxygraph-2k Manual Titrations: MiPNet09.12
- Titration volume: 1 µl steps using a 10 µl syringe
- Final conc. in 2 ml O2k-chamber: 0.5 µM steps
Frequently, more diluted FCCP for getting several titration steps is necessary. In this case prepare more diluted solution (ditution 2-10x) from your stock solution.