Difference between revisions of "SUIT-018 AmR mt D041"
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::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(GM) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(GM) | ||
::: '''[[SUIT protocol pattern]]:''' lineal 1GMS;2D;3Ama | ::: '''[[SUIT protocol pattern]]:''' lineal 1GMS;2D;3Ama | ||
SUIT-018 AmR mt D041 is a short protocol to study simultaneously the oxygen dependence of O<sub>2</sub> flux and H<sub>2</sub>O<sub>2</sub> production on isolated mitochondria or tissue homogenate (except of liver homogenate). During the development of this protocol the [O<sub>2</sub>] will be modify | SUIT-018 AmR mt D041 is a short protocol to study simultaneously the oxygen dependence of O<sub>2</sub> flux and H<sub>2</sub>O<sub>2</sub> production on isolated mitochondria or tissue homogenate (except of liver homogenate). During the development of this protocol the [O<sub>2</sub>] will be modify injecting nitrogen in the chamber. | ||
__TOC__ | __TOC__ |
Revision as of 17:21, 12 February 2019
Description
Abbreviation: NS(GM)
Reference: A: SUIT-018 simultaneous determination of the oxygen dependence of O2 flux and H2O2 production flux on isolated mitochondria and tissue homogenate at low oxygen concentration modifying the oxygen concentration in the same protocol.
SUIT number: D041_1GMS;2D;3Ama
O2k-Application: AmR
- SUIT-category: NS(GM)
- SUIT protocol pattern: lineal 1GMS;2D;3Ama
SUIT-018 AmR mt D041 is a short protocol to study simultaneously the oxygen dependence of O2 flux and H2O2 production on isolated mitochondria or tissue homogenate (except of liver homogenate). During the development of this protocol the [O2] will be modify injecting nitrogen in the chamber.
Communicated by Iglesias-Gonzalez J and Komlodi T (last update 2019-02-12)
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
MiPNet24.10 H2O2 flux analysis | 2021-10-22 | Hydrogen peroxide flux analysis using Amplex UltraRed assay in MiR05-Kit with DatLab 7.4 | ||
Komlodi 2021 BEC AmR-O2 | 2021 | Komlรณdi T, Sobotka O, Gnaiger E (2021) Facts and artefacts on the oxygen dependence of hydrogen peroxide production using Amplex UltraRed. Bioenerg Commun 2021.4. https://doi.org/10.26124/bec:2021-0004 | Saccharomyces cerevisiae | Other cell lines |
MiPNet20.14 AmplexRed H2O2-production | 2019-06-24 | O2k-FluoRespirometry: HRR and simultaneous determination of H2O2 production with Amplex UltraRed. | Mouse | Heart |
Komlodi 2018 Methods Mol Biol | 2018 | Komlodi T, Sobotka O, Krumschnabel G, Bezuidenhout N, Hiller E, Doerrier C, Gnaiger E (2018) Comparison of mitochondrial incubation media for measurement of respiration and hydrogen peroxide production. Methods Mol Biol 1782:137-55. | Human Mouse | Skeletal muscle HEK |
Makrecka-Kuka 2015 Biomolecules | 2015 | 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. https://doi.org/10.3390/biom5031319 | Human Mouse | Heart Nervous system HEK |
Krumschnabel 2015 Methods Mol Biol | 2015 | 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. | Mouse | Nervous system |
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
0DTPA |
| |||
0SOD |
| |||
0HRP |
| |||
0AmR |
|
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1GMS | GMSL(n) | NS | CI&CII | 1GMS
|
2D | GMSP | NS | CI&CII | 1GMS;2D
|
3Ama | ROX | 1GMS;2D;3Ama
|
Strengths and limitations
- + Reasonable duration of the experiment.
- + Simple protocol to evaluate the oxygen dependence of H2O2 production in LEAK and OXPHOS.
- - Measurements with Amplex UltraRed assay cannot be carried out with liver homogenate.
- - Reoxygenation and nitrogen injection could lead to bubble formation in the O2k-Chamber
- - CIV activity and cytochrome c test cannot be performed together with the fluorescence.
Compare SUIT protocols
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Fluorometry