Difference between revisions of "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 (2018-11-26) Mitochondr Physiol Network

Abstract: Krumschnabel G, Fontana-Ayoub M, Fasching M, Gnaiger E (2018) O2k-FluoRespirometry: HRR and simultaneous determination of H₂O₂ production with Amplex UltraRed. Mitochondr Physiol Network 20.14(03):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:

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

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

Coupling state: LEAK, OXPHOS, ET Pathway: S 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

Template:Keywords H2O2

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 {{Publication
-|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-Fluorometry: HRFR and simultaneous determination of H<sub>2</sub>O<sub>2</sub> production with Amplex UltraRed.
+|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-FluoRespirometry: HRR and simultaneous determination of H<sub>2</sub>O<sub>2</sub> production with Amplex UltraRed.
 |info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/5/57/MiPNet20.14_AmplexRed_H2O2-production.pdf|Bioblast pdf]] »[http://wiki.oroboros.at/index.php/File:MiPNet20.14_AmplexRed_H2O2-production.pdf Versions]
 |authors=Oroboros
 |year=2018-11-26
 |journal=Mitochondr Physiol Network
-|abstract='''Krumschnabel G, Fontana-Ayoub M, Fasching M, Gnaiger E (2018) O2k-Fluorometry: HRFR and simultaneous determination of H<sub>2</sub>O<sub>2</sub> production with Amplex UltraRed. Mitochondr Physiol Network 20.14(03):1-6.'''
+|abstract='''Krumschnabel G, Fontana-Ayoub M, Fasching M, Gnaiger E (2018) O2k-FluoRespirometry: HRR and simultaneous determination of H<sub>2</sub>O<sub>2</sub> production with Amplex UltraRed. Mitochondr Physiol Network 20.14(03):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: [[O2k-Publications:_O2k-Fluorometry_-_Fluorophores|H<sub>2</sub>O<sub>2</sub> production, mitochondrial membrane potential, intracellular pH, Ca<sup>2+</sup>, Mg<sup>2+</sup>, and NADH levels]].