FCCP: Difference between revisions
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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. | |||
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'''Preparation of 1 mM stock solution''' (dissolved in 99.9% | '''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. | ||
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'''Oxygraph-2k | '''Oxygraph-2k manual titrations:''' [[MiPNet09.12]] | ||
::* Titration volume: 1 µl steps using a 10 µl syringe | ::* Titration volume: 1 µl steps using a 10 µl syringe | ||
Revision as of 21:34, 10 January 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: MiPNet10.04; SteinlechnerMaran_1996_AJP
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).
- FCCP should be used instead of CCCP at low experimental temperature, or in laboratories at low room temperature that may prevail during expeditions, to avoid blocking the needle of the Hamilton titration syringe.
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!
Caution: Chemicals stored in the fridge or freezer should be allowed to reach room temperature before opening.
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 stock solution.
- 3) Store in glass vials (recommendation) at -20 °C.
Oxygraph-2k manual titrations: MiPNet09.12
- Titration volume: 1 µl steps using a 10 µl syringe
- 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.
