MiPNet20.14 AmplexRed H2O2-production

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O2k-FluoRespirometry: HRR and simultaneous determination of H2O2 production with Amplex UltraRed.

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Oroboros (2019-06-24) Mitochondr Physiol Network

Abstract: Krumschnabel G, Fontana-Ayoub M, Fasching M, Gnaiger E (2019) O2k-FluoRespirometry: HRR and simultaneous determination of H2O2 production with Amplex UltraRed. Mitochondr Physiol Network 20.14(04):1-6.

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:

  1. Quality control of preparations by respirometric performance.
  2. Objective exclusion criteria for protocol related problems, e.g. overtitration of uncouplers, side effects of TPP+ or fluorophores.
  3. Elimination of artifacts of normalization for a mitochondrial marker when combining two methods in a single chamber.
  4. Additional information is acquired for a limited amount of sample.
  5. 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:

O2k-Publications in the MiPMap 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«

O2k-Publications in the MiPMap

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 


Organism: Mouse  Tissue;cell: Heart  Preparation: Isolated mitochondria 


Coupling state: LEAK, OXPHOS, ET  Pathway:HRR: Oxygraph-2k, O2k-Fluorometer, O2k-Protocol 

O2k-Demo, 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 


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» Keywords: Hydrogen peroxide