Difference between revisions of "FCCP"
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'''Preparation of 1 mM stock solution''' (dissolved in 99.9% Ethanol): | '''Preparation of 1 mM stock solution''' (dissolved in 99.9% Ethanol): | ||
::1) weigh 1.27 mg of FCCP into a glass vial | ::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 | ::3) store in glass vials (recommendation) at -20 °C | ||
'''Oxygraph-2k Manual Titrations:''' [[MiPNet09.12]] | '''Oxygraph-2k Manual Titrations:''' [[MiPNet09.12]] |
Revision as of 11:23, 18 May 2012
Description
FCCP (synonym:Carbonyl cyanide p-(trifluoro-methoxy) phenyl-hydrazone) uncoupler, Fc: FCCP added at concentration c; c is the optimum uncoupler concentration in titrations of FCCP to obtain maximum mitochondrial respiration in the noncoupled state of ETS capacity.
Abbreviation: Fc
Reference: MiPNet10.04; SteinlechnerMaran_1996_AJP
MitoPedia topics: Uncoupler
Application in HRR
F: FCCP (Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, C10H5F3N4O); Sigma C 2920, 10 mg, store at 4 °C: FW = 254.17)
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.