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Difference between revisions of "FCCP"

From Bioblast
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== CCCP versus FCCP ==
== CCCP versus FCCP ==
'''[[Bioblast_alert#Bioblast_alert_2013.2802.29:_2013-08-08|Bioblast alert 2013(02)]]''': We replace FCCP by the less expensive '''[[CCCP]]''' in our experiments with high-resultion respirometry ([[HRR]]).
'''[[Bioblast_alert#Bioblast_alert_2013.2802.29:_2013-08-08|Bioblast alert 2013(02)]]''': We replace FCCP by the less expensive '''[[CCCP]]''' in our experiments with high-resultion respirometry ([[HRR]]).
* FCCP should be used instead of CCCP at [[O2k-Publications: Temperature|low experimental temperature]], or in laboratories at low room temperature that may prevail during [http://www.oroboros.at/?expeditions expeditions], to avoid blocking the needle of the Hamilton titration syringe.





Revision as of 05:13, 22 April 2014


high-resolution terminology - matching measurements at high-resolution


FCCP

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: Steinlechner-Maran 1996 Am J Physiol Cell Physiol, Gnaiger_2008_POS



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!


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 °C.

Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.

Oxygraph-2k manual titrations MiPNet09.12 O2k-Titrations

  • Titration volume: 1 µl (or 0.5 µl) steps using a 10 µl syringe (2 ml O2k-Chamber).
  • Final concentration: 0.5 µM (or 0.25 µM) 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]

  1. 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: 327-352. »Bioblast Access