FCCP: Difference between revisions
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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 | ::1) weigh 1.27 mg of FCCP into a glass vial and dissolve in 5 ml Ethanol | ||
::2) 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]] | '''Oxygraph-2k Manual Titrations:''' [[MiPNet09.12]] | ||
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::* Titration volume: 1 µl steps using a 10 µl syringe | ::* Titration volume: 1 µl steps using a 10 µl syringe | ||
::* Final conc. in 2 ml O2k-chamber: 0.5 µM steps | ::* 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. | |||
Revision as of 11:22, 7 December 2010
- high-resolution terminology - matching measurements at high-resolution
FCCP
Description
Carbonyl cyanide p-(trifluoro-methoxy) phenyl-hydrazone (C10H5F3N4O); uncoupler, F(x): FCCP added at concentration x.
Abbreviation: Fx
Reference: MiPNet10.04; SteinlechnerMaran_1996_AJP
Labels:
Regulation: Respiration; OXPHOS; ETS Capacity"Respiration; OXPHOS; ETS Capacity" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.
HRR: Chemicals; Media"Chemicals; Media" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property.
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 and dissolve in 5 ml Ethanol
- 2) 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.
