Difference between revisions of "FCCP"
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{{MitoPedia | {{MitoPedia | ||
|abbr=FCCP | |abbr=FCCP | ||
|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]]: | |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 [[ETS 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 == | ||
'''[[Bioblast_alert#Bioblast_alert_2013.2802.29:_2013-08-08|Bioblast alert 2013(02)]]''': We replace FCCP by the less expensive and equally effective '''[[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 and equally effective '''[[CCCP]]''' in our experiments with high-resultion respirometry ([[HRR]]). | ||
== Application in HRR == | == Application in HRR == | ||
''' | :::: '''FCCP''' (Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, C<sub>10</sub>H<sub>5</sub>F<sub>3</sub>N<sub>4</sub>O); Sigma C 2920, 10 mg, store at 4 °C: FW = 254.17) | ||
<span style="color:#8B008B"> '''Caution:''' Toxic!</span> | :::: <span style="color:#8B008B"> '''Caution:''' Toxic!</span> | ||
'''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. | ::::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. | ::::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. | ::::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. | :::: '''Caution:''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening. | ||
'''O2k manual titrations''' [[MiPNet09.12 O2k-Titrations]] | ::: '''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). | ::::* 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. | ::::* 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. <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> | ||
<references/> | <references/> |
Revision as of 06:21, 25 October 2016
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 ETS 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]
- ↑ 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«