Difference between revisions of "FCCP"
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|description='''FCCP''' (Carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone, C<sub>10</sub>H<sub>5</sub>F<sub>3</sub>N<sub>4</sub>O) is a protonophore or [[uncoupler]]: added at uncoupler concentration U<sub>''c''</sub>; ''c'' is the [[optimum uncoupler concentration]] in titrations to obtain maximum mitochondrial respiration in the [[noncoupled respiration|noncoupled]] state of [[ET-capacity]]. | |description='''FCCP''' (Carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone, C<sub>10</sub>H<sub>5</sub>F<sub>3</sub>N<sub>4</sub>O) is a protonophore or [[uncoupler]]: added at uncoupler concentration U<sub>''c''</sub>; ''c'' is the [[optimum uncoupler concentration]] in titrations to obtain maximum mitochondrial respiration in the [[noncoupled respiration|noncoupled]] state of [[ET-capacity]]. | ||
|info=[[Steinlechner-Maran 1996 Am J Physiol Cell Physiol]], [[Gnaiger_2008_POS]] | |info=[[Steinlechner-Maran 1996 Am J Physiol Cell Physiol]], [[Gnaiger_2008_POS]] | ||
}} | }} | ||
== CCCP versus FCCP == | == CCCP versus FCCP == | ||
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:::: 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. <ref> 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:327-52. [[Gnaiger_2008_POS |»Bioblast link«]]</ref> | :::: 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. <ref> 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:327-52. [[Gnaiger_2008_POS |»Bioblast link«]]</ref> | ||
{{Keywords: Uncoupling}} | |||
<references/> | <references/> | ||
{{MitoPedia topics | |||
|mitopedia topic=Uncoupler | |||
}} | |||
Revision as of 07:15, 19 February 2020
- high-resolution terminology - matching measurements at high-resolution
FCCP
Description
FCCP (Carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone, C10H5F3N4O) is a protonophore or uncoupler: added at uncoupler concentration Uc; c is the optimum uncoupler concentration in titrations to obtain maximum mitochondrial respiration in the noncoupled state of ET-capacity.
Abbreviation: FCCP
Reference: Steinlechner-Maran 1996 Am J Physiol Cell Physiol, Gnaiger_2008_POS
CCCP versus FCCP
- Bioblast alert 2013(02): We replace FCCP by the less expensive and equally effective CCCP in our experiments with high-resultion respirometry (HRR).
Application in HRR
- 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.
- O2k 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]
- Bioblast links: Uncoupling - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Specific
- » Artefacts by single dose uncoupling
- » ATP synthase
- » CCCP
- » Coupling-control protocol
- » DNP
- » Dyscoupled respiration
- » FCCP
- » Is respiration uncoupled - noncoupled - dyscoupled?
- » Noncoupled respiration: Discussion
- » Uncoupler
- » Uncoupled respiration - see » Noncoupled respiration
- » Uncoupling proteins
- » Uncoupling protein 1
- » Uncoupler titrations - Optimum uncoupler concentration
- Specific
- Respiratory states and control ratios
- » Biochemical coupling efficiency
- » Coupling-control state
- » Electron-transfer-pathway state
- » Electron-transfer pathway
ET capacity- » E-L coupling efficiency
- » Flux control efficiency
- » Flux control ratio
- » LEAK-control ratio
- » LEAK respiration
- » Noncoupled respiration
- » OXPHOS
- » OXPHOS capacity; » State 3
- » OXPHOS-control ratio, P/E ratio
- » Respiratory acceptor control ratio
- » ROUTINE-control ratio
- » ROUTINE respiration
- » ROUTINE state
- » State 3u
- » State 4
- » Uncoupling-control ratio UCR
- Respiratory states and control ratios
- Gnaiger E et al ― MitoEAGLE Task Group (2020) Mitochondrial physiology. Bioenerg Commun 2020.1. https://doi.org/10.26124/bec:2020-0001.v1
- Gnaiger E (2020) Mitochondrial pathways and respiratory control. An introduction to OXPHOS analysis. 5th ed. Bioenerg Commun 2020.2. https://doi.org/10.26124/bec:2020-0002
- General (alphabetical order)
- Other keyword lists
- ↑ 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:327-52. »Bioblast link«
MitoPedia topics:
Uncoupler
