Difference between revisions of "MiPNet03.02 Chemicals-Media"

Latest revision as of 17:38, 17 October 2023

MiPNet03.02 Chemicals-Media

Selected media and chemicals for respirometry with mitochondrial preparations.

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Oroboros (2016-08-30) Mitochondr Physiol Network

Abstract: Fontana-Ayoub M, Fasching M, Gnaiger E (2016) Selected media and chemicals for respirometry with mitochondrial preparations. Mitochondr Physiol Network 03.02(18):1-10. Different media for tissue preparation and respiration are used in investigations of mitochondrial function. Initial decisions on the composition of media and chemicals are decisive for long-term studies and crucial for comparability of results. As a guideline, we summarize an update of our experience with media and chemicals for high-resolution respirometry with isolated mitochondria, permeabilized cells, muscle fibres and tissue homogenates. Whereas optimization is necessary for specific experimental protocols, standardization will improve the comparability of results obtained in different laboratories. Efforts towards standardization are important for the advancement of mitochondrial physiology.

» Product: Oroboros O2k, O2k-Catalogue

• O2k-Network Lab: AT Innsbruck Oroboros

Supplementary information

» MiPNet09.12 O2k-Titrations; MitoPedia

» Cytochrome c control factor

» For calculations, see Excel file: O2k-Titrations.xls

» For pH adjustment of BIOPS solution, note the temperature dependence:

at 0 °C pH = 7.1
at 23 °C pH = 6.75

Cited by

Cardoso et al (2021) Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration. Bioenerg Commun 2021.1. doi:10.26124/bec:2021-0001
Komlodi et al (2022) Hydrogen peroxide production, mitochondrial membrane potential and the coenzyme Q redox state measured at tissue normoxia and experimental hyperoxia in heart mitochondria. MitoFit Preprints 2021 (in prep)

Labels: MiParea: Instruments;methods

HRR: O2k-Protocol

O2k-chemicals and media, Uncoupling, ADP, ATP, MitoFit 2021 MgG, MitoFit 2021 Tissue normoxia, MitoFit 2022 NADH

@@ Line 1: / Line 1: @@
+{{OROBOROS header page name}}
 {{Publication
-|title=Fontana-Ayoub M, Eigentler A, Fasching M, Gnaiger E (2013) Selected media and chemicals for respirometry with mitochondrial preparations. Mitochondr Physiol Network 03.02.
+|title=Selected media and chemicals for respirometry with mitochondrial preparations. [[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]]
-|info=[[Media:MiPNet03.02 Chemicals-Media.pdf| '''Open Access-MiPNet03.02.pdf''']], [http://www.bioblast.at/index.php/File:MiPNet03.02_Chemicals-Media.pdf Versions] [[Image:O2k-Protocols.jpg|right|150px|link=http://www.oroboros.at/?O2k-protocols|O2k-Protocols contents]]
+|info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/3/3c/MiPNet03.02_Chemicals-Media.pdf |Bioblast pdf]]  »[http://www.bioblast.at/index.php/File:MiPNet03.02_Chemicals-Media.pdf Versions]
-|authors=OROBOROS
+|authors=Oroboros
-|year=2013
+|year=2016-08-30
 |journal=Mitochondr Physiol Network
-|abstract=Different media for tissue preparation and respiration are used in investigations of mitochondrial function. Initial decisions on the composition of media and chemicals are decisive for long-term studies and crucial for comparability of results. As a guideline, we summarize an update of our experience with media and chemicals for high-resolution respirometry with isolated mitochondria, permeabilized cells, muscle fibres and tissue homogenates. Whereas optimization is necessary for specific experimental protocols, standardization will improve the comparability of results obtained in different laboratories. Efforts towards standardization are important for the advancement of mitochondrial physiology.
+|abstract='''Fontana-Ayoub M, Fasching M, Gnaiger E (2016) Selected media and chemicals for respirometry with mitochondrial preparations. Mitochondr Physiol Network 03.02(18):1-10.'''
+Different media for tissue preparation and respiration are used in investigations of mitochondrial function. Initial decisions on the composition of media and chemicals are decisive for long-term studies and crucial for comparability of results. As a guideline, we summarize an update of our experience with media and chemicals for high-resolution respirometry with isolated mitochondria, permeabilized cells, muscle fibres and tissue homogenates. Whereas optimization is necessary for specific experimental protocols, standardization will improve the comparability of results obtained in different laboratories. Efforts towards standardization are important for the advancement of mitochondrial physiology.
+:» Product: [[Oroboros O2k]], [[Oroboros O2k-Catalogue | O2k-Catalogue]]
+|mipnetlab=AT Innsbruck Oroboros
+}}
+== Supplementary information ==
+::::» [[MiPNet09.12 O2k-Titrations]]; [[MitoPedia]]
+::::» [[Cytochrome c control factor]]
+::::» For calculations, see Excel file: [http://www.oroboros.at/?Protocols_titrations O2k-Titrations.xls]
+::::» For pH adjustment of BIOPS solution, note the temperature dependence:
+:::::::* at    0 °C pH = 7.1
+:::::::* at   23 °C pH = 6.75
+== Cited by ==
+{{Template:Cited by Cardoso 2021 MitoFit MgG}}
+{{Template:Cited by Komlodi 2021 MitoFit Tissue normoxia}}
-:>> O2k-Protocols:[http://www.oroboros.at/?o2k-protocols Overall contents]
-:>> Product: [http://www.oroboros.at/?oxygraph OROBOROS Oxygraph-2k], [[O2k-Catalogue_OROBOROS| O2k-Catalogue]]
-|mipnetlab=AT Innsbruck OROBOROS,
-|discipline=Mitochondrial Physiology
-}}
 {{Labeling
 |area=Instruments;methods
-|instruments=Protocol
+|instruments=O2k-Protocol
-|additional=Chemicals & Media
+|additional=O2k-chemicals and media, Uncoupling, ADP, ATP, MitoFit 2021 MgG, MitoFit 2021 Tissue normoxia, MitoFit 2022 NADH
-|discipline=Mitochondrial Physiology
 }}
-== Further information ==
-:>> [[MiPNet09.12 O2k-Titrations]]; [[MitoPedia Glossaries]]
-:>> [http://www.oroboros.at/?CytochromecControl Cytochrome ''c'' control]
-:>> For calculations, see Excel file: [http://www.oroboros.at/?Protocols_titrations O2k-Titrations.xls]
-== History ==
-* Malate was listed for a final concentration of 2 mM until 2013. During this year, test exeriments with mitochondria from various tissues and species and with different mt-preparations (isolated mitochondria, permeabilized fibres, tissue homogenate) revealed an inhibitory effect of 2 mM malate on CII linked respiration (S+Rot). The inhibitory effect is less at 0.5 mM malate, and 0.5 mM malate may be saturating for CI-linked respiration. Evaluation is required for the optimum malate concentrations in SUIT protocols with sequential CI,CI+II and CII substrate states.
-* [[Carbonyl cyanide m-chloro phenyl hydrazone|From FCCP to CCCP]]
-* Up to 2006, rotenone was added at a high final concentration (0.5 µM), hence a 0.5 mM stock solution was prepared. Since 0.05 µM may be fully inhibiting, we suggested to use a stock solution at lower concentration (0.1 mM), to reduce the problem of rotenone retention. The 2006 edition listed a 0.2 mM stock solution, but the amount added to 10 ml ethanol was given for a 0.15 mM stock solution. Our recent rotenone titrations with permeabilized muscle fibers, however, confirmed that the originally proposed higher rotenone concentration is actually required for full inhibition.