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Difference between revisions of "FCCP"

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{{MitoPedia
{{MitoPedia
|abbr=U<sub>''c''</sub>
|abbr=FCCP
|description='''FCCP''' (Carbonyl cyanide p-trifluoro-methoxyphenyl hydrazone) 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 [[ET capacity]].
|info=[[MiPNet10.04]]; [[SteinlechnerMaran_1996_AJP]]
|info=[[Steinlechner-Maran 1996 Am J Physiol Cell Physiol]], [[Gnaiger_2008_POS]]
|type=Uncoupler
}}
{{MitoPedia methods|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 '''[[CCCP]]''' in our experiments with high-resultion respirometry ([[HRR]]).
:::: '''[[Bioblast_alert_2013#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]] ==
 
:::: '''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: M = 254.17 g·mol<sup>-1</sup>)
:::: Ethanol stock solutions may be stored in 5 ml glass vials (instead of freezing 0.2 mL portions in small vials). Use subsamples during experiments to avoid contamination of the storage solution. Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.
:::: <span style="color:#8B008B"> '''Caution:''' Toxic!</span>
 
 
::: '''Preparation of 1 mM stock solution''' (dissolved in 99.9% ethanol) for '''2-mL O2k-chamber''':


::::# Weigh 1.27 mg of FCCP into a glass vial.
::::# Dissolve in 5 ml ethanol.
::::# Divide into 0.2 ml portions.
::::# Store in glass vials at -20 °C.


== Application in HRR ==
::: '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]


'''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)
::::* Titration volume ('''2-mL O2k-chamber'''): 1 µl (or 0.5 µl) steps using a 10 µl syringe.
::::* Final concentration: 0.5 µM (or 0.25 µM) steps.




<span style="color:#8B008B"> '''Caution:''' Toxic!</span>
:::: '''Preparation of 0.25 mM stock solution''' (dissolved in Ethanol) for '''0.5-mL O2k-chamber''':


<span style="color:#8B008B"> '''Caution:''' Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.</span>
::::# Take 100 µL of 1 mM stock solution.
::::# Dilute with 300 µL ethanol.


:::» '''O2k manual titrations'''  [[MiPNet09.12 O2k-Titrations]]


'''Preparation of 1 mM stock solution''' (dissolved in 99.9% Ethanol):
::::* Titration volume ('''0.5-mL O2k-chamber'''): 1 µL (or 0.5 µL) steps using a 10 µL Hamilton syringe.
::::* Final concentration: 0.5 µM (or 0.25 µM) steps.


::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 stock solution.
::3) Store in glass vials (recommendation) at -20 °C.


:::: 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>


'''Oxygraph-2k Manual Titrations:'''  [[MiPNet09.12]]
{{Keywords: Uncoupling}}


::* Titration volume: 1 µl steps using a 10 µl syringe
<references/>
::* 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.
{{MitoPedia topics
|mitopedia topic=Uncoupler
}}

Revision as of 09:29, 22 March 2021


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: M = 254.17 g·mol-1)
Ethanol stock solutions may be stored in 5 ml glass vials (instead of freezing 0.2 mL portions in small vials). Use subsamples during experiments to avoid contamination of the storage solution. Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.
Caution: Toxic!


Preparation of 1 mM stock solution (dissolved in 99.9% ethanol) for 2-mL O2k-chamber:
  1. Weigh 1.27 mg of FCCP into a glass vial.
  2. Dissolve in 5 ml ethanol.
  3. Divide into 0.2 ml portions.
  4. Store in glass vials at -20 °C.
O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (2-mL O2k-chamber): 1 µl (or 0.5 µl) steps using a 10 µl syringe.
  • Final concentration: 0.5 µM (or 0.25 µM) steps.


Preparation of 0.25 mM stock solution (dissolved in Ethanol) for 0.5-mL O2k-chamber:
  1. Take 100 µL of 1 mM stock solution.
  2. Dilute with 300 µL ethanol.
» O2k manual titrations MiPNet09.12 O2k-Titrations
  • Titration volume (0.5-mL O2k-chamber): 1 µL (or 0.5 µL) steps using a 10 µL Hamilton syringe.
  • 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]


Questions.jpg


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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
Respiratory states and control ratios
» Biochemical coupling efficiency
» Coupling-control state
» Electron-transfer-pathway state
» Electron-transfer pathway
E.jpg 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
General (alphabetical order)
» Adenine nucleotide translocase
» Adenylates
» Electron transfer pathway
» Mitochondrial preparations
» mt-membrane potential
» Oxygen flux
» Phosphorylation system
» Proton leak
» Proton slip
» TIP2k
Other keyword lists
» Template:Keywords: Force and membrane potential


  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«


MitoPedia topics: Uncoupler