Difference between revisions of "MiPNet24.10 H2O2 flux analysis"
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|area=Respiration, Instruments;methods | |area=Respiration, Instruments;methods | ||
|instruments=Oxygraph-2k, O2k-Fluorometer, O2k-Protocol | |instruments=Oxygraph-2k, O2k-Fluorometer, O2k-Protocol | ||
|additional= | |additional=O2k-MultiSensor, MitoFitPublication, SUIT-013, SUIT-013 AmR ce D023, SUIT-009, SUIT-009 AmR mt D021, SUIT-009 AmR pce D019, SUIT-018, SUIT-018 AmR mt D031, SUIT-006 AmR mt D048, SUIT-003 AmR ce D017, SUIT-003 AmR ce D058, O2k-Analysis, DatLab, | ||
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Revision as of 16:06, 9 October 2019
MiPNet24.10 H2O2 flux analysis
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Oroboros (2019-08-05) Mitochondr Physiol Network
Abstract: Komlodi T, Iglesias-Gonzalez J, Cardoso LHD, Sobotka O, Gnaiger E (2019) Hydrogen peroxide flux analysis using Amplex UltraRed assay in MiR05-Kit with DatLab 7.4. Mitochondr Physiol Network 24.10(01):1-5.
High-resolution respirometry is extended by O2k-MultiSensor modules allowing the simultaneous measurement of oxygen consumption and one additional parameter. Fluorometric methods are available for a wide range of analytical parameters of major interest in mitochochondrial physiology: H2O2 production, mitochondrial membrane potential, intracellular pH, Ca2+, Mg2+, and NADH levels. Advantages of the simultaneous measurement of additional parameters in a single respirometric chamber:
- Quality control of preparations by respirometric performance.
- Objective exclusion criteria for protocol related problems, e.g. overtitration of uncouplers, side effects of TPP+ or fluorophores.
- Elimination of artifacts of normalization for a mitochondrial marker when combining two methods in a single chamber.
- Additional information is acquired for a limited amount of sample.
- Oxygen dependence of parameters can be studied.
In the publication by Krumschnabel et al. 2015 we introduce the O2k-Fluo LED2-Module, describe one selected stage in its development, and show its application for the simultaneous measurement of respiration and H2O2 production of mouse brain mitochondria. The paper by Makrecka-Kuka et al. 2015 focuses on the application of the Amplex red assay for determining H2O2 production in three different models of mitochondrial preparations.
O2k-Publication:
Krumschnabel G, Fontana-Ayoub M, Sumbalova Z, Heidler J, Gauper K, Fasching M, Gnaiger E (2015) Simultaneous high-resolution measurement of mitochondrial respiration and hydrogen peroxide production. Methods Mol Biol 1264:245-61. »Bioblast link«
Makrecka-Kuka M, Krumschnabel G, Gnaiger E (2015) High-resolution respirometry for simultaneous measurement of oxygen and hydrogen peroxide fluxes in permeabilized cells, tissue homogenate and isolated mitochondria. Biomolecules 5:1319-38. »Bioblast link«
• O2k-Network Lab: AT_Innsbruck_Oroboros
Labels: MiParea: Respiration, Instruments;methods
HRR: Oxygraph-2k, O2k-Fluorometer, O2k-Protocol
O2k-MultiSensor, MitoFitPublication, SUIT-013, SUIT-013 AmR ce D023, SUIT-009, SUIT-009 AmR mt D021, SUIT-009 AmR pce D019, SUIT-018, SUIT-018 AmR mt D031, SUIT-006 AmR mt D048, SUIT-003 AmR ce D017, SUIT-003 AmR ce D058, O2k-Analysis, DatLab
