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Difference between revisions of "FCCP"

From Bioblast
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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 directly into a 5 ml glass vial.
::1) weigh 1.27 mg of FCCP into a glass vial and dissolve in 5 ml Ethanol
::2) add 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) 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
::4) 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.