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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]]: '''FCCP''' 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]].
|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]]
|type=Uncoupler
}}
{{MitoPedia concepts|type=Uncoupler
}}
{{MitoPedia methods|type=Uncoupler
}}
{{MitoPedia O2k and high-resolution respirometry|type=Uncoupler
}}
{{MitoPedia topics
|mitopedia topic=Uncoupler
|type=Uncoupler
}}
}}
== 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 ==


'''F: 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)
:::: '''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


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 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]
  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«