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Difference between revisions of "MiPNet24.10 H2O2 flux analysis"

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{{OROBOROS header page name}}
{{Publication
{{Publication
|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] Hydrogen peroxide flux analysis using Amplex UltraRed assay in MiR05-Kit with DatLab 7.4
|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] Hydrogen peroxide flux analysis using Amplex UltraRed assay in MiR05-Kit with DatLab 7.4
|info=[[File:PDF.jpg|100px|link=https://wiki.oroboros.at/images/b/ba/MiPNet24.10_H2O2_analysis_template.pdf|Bioblast pdf]] »[http://wiki.oroboros.at/index.php/File:MiPNet20.14_AmplexRed_H2O2-production.pdf Versions]
|info=[[File:PDF.jpg|100px|link=https://wiki.oroboros.at/images/b/ba/MiPNet24.10_H2O2_analysis_template.pdf|Bioblast pdf]] »[https://www.bioblast.at/index.php/File:MiPNet24.10_H2O2_analysis_template.pdf Versions]
|authors=Oroboros
|authors=Oroboros
|year=2019-08-05
|year=2021-10-22
|journal=Mitochondr Physiol Network
|journal=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.'''
|abstract=Komlodi T, Cardoso LHD, Gnaiger E (2021) Hydrogen peroxide flux analysis using Amplex UltraRed assay in MiR05-Kit with DatLab 7.4. Mitochondr Physiol Network 24.10(03):1-5.  
 
{{MiPNet pdf page linking to MitoPedia}}
[[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]].
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<sup>+</sup> 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_2015_Methods_Mol_Biol|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 H<sub>2</sub>O<sub>2</sub> production of mouse brain mitochondria. The paper by [[Makrecka-Kuka_2015_Biomolecules|Makrecka-Kuka et al. 2015]] focuses on the application of the Amplex red assay for determining H<sub>2</sub>O<sub>2</sub> production in three different models of mitochondrial preparations.
 
 
'''O2k-Publication''':
[[Image:O2k-Publications.jpg|50px|link=O2k-Publications:_Topics|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. [[Krumschnabel 2015 Methods Mol Biol |»Bioblast link«]]
 
[[Image:O2k-Publications.jpg|50px|left|link=O2k-Publications:_Topics|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. [[Makrecka-Kuka 2015 Biomolecules| »Bioblast link«]]
|mipnetlab=AT_Innsbruck_Oroboros
|mipnetlab=AT_Innsbruck_Oroboros
}}
}}
'''Acknowledgements'''
[[File:Template NextGen-O2k.jpg|left|400px|link=NextGen-O2k]]
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{{Labeling
{{Labeling
|area=Respiration, Instruments;methods
|area=Respiration, Instruments;methods
|instruments=Oxygraph-2k, O2k-Fluorometer, O2k-Protocol
|instruments=Oxygraph-2k, O2k-Fluorometer, O2k-Protocol
|additional=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, O2k-Analysis, DatLab,
|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, DL7.4
}}
}}
{{OROBOROS header page name}}
{{Keywords H2O2}}

Latest revision as of 08:55, 22 October 2021

                



MiPNet24.10 H2O2 flux analysis

Publications in the MiPMap
O2k-Protocols
Hydrogen peroxide flux analysis using Amplex UltraRed assay in MiR05-Kit with DatLab 7.4

» Bioblast pdf »Versions

Oroboros (2021-10-22) Mitochondr Physiol Network

Abstract: Komlodi T, Cardoso LHD, Gnaiger E (2021) Hydrogen peroxide flux analysis using Amplex UltraRed assay in MiR05-Kit with DatLab 7.4. Mitochondr Physiol Network 24.10(03):1-5.

O2k-technical support and open innovation
Open the pdf document above.
» Current O2k-series: NextGen-O2k Series XB and O2k Series J
» Current software versions DatLab 8.0: MitoPedia: DatLab


O2k-Network Lab: AT_Innsbruck_Oroboros

Acknowledgements

Template NextGen-O2k.jpg






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, DL7.4