Difference between revisions of "MiPNet21.06 SUIT RP"
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| [[Media:SUIT_RP1- | | [[Media:SUIT_RP1-mt.docx |»SUIT_RP1mt]] | ||
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| [[Media:SUIT_RP2- | | [[Media:SUIT_RP2-mt.docx |»SUIT_RP2mt]] | ||
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| [[Media:RP print.pdf |»RP print]] | | [[Media:RP print.pdf |»RP print]] |
Revision as of 14:41, 1 July 2016
SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. |
OROBOROS (2016-01-23) Mitochondr Physiol Network
Abstract: Doerrier C, Sumbalova Z, Lamberti G, Krumschnabel G, Gnaiger (2016) SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. Mitochondr Physiol Network 21.06(pre03):1-10. » Versions
- » MitoPedia: SUIT reference protocol
- » Instrument: O2k, O2k-Catalogue
• O2k-Network Lab: AT_Innsbruck_OROBOROS
Labels: MiParea: Instruments;methods
Preparation: Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria
Regulation: Substrate Coupling state: LEAK, OXPHOS, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property.
HRR: Oxygraph-2k, O2k-Protocol
MitoPathways, MitoFitPublication
Work in progress
RP
- RP1: Compared to RP1-T01, move Oct titration after S.
- RP2: Compared to RP2-T01, move U titration after Gp.
RP-T01
Cytochrome c test
- In protocol RP2, the cytochrome c test might better be moved from a c-titration after pyruvate (P) to a titration after M2. Otherwise, there is a risk of underestimation of FAO in cases of a cytochrome c effect. On the other hand, at the low flux before titration of P, the sensitivity of the cytochrome c test may be low.
- ~ Gnaiger Erich 18:26, 23 January 2016 (CET)
Mark names
- Even the highly abbreviated names of respiratory states have become too long for DatLab, where mark names are restricted to 8 digits. The visibility in DatLab of long mark names is restricted. The simplest solution for short mark names is the use of a numerical sequence, with the preceeding event name added for information.
- ~ Gnaiger Erich 16:50, 23 January 2016 (CET)
Laboratory sheets for SUIT protocols RP1 and RP2
Type | SUIT protocol | Sample | Abbr | docx file | pdf file | Last update |
---|---|---|---|---|---|---|
Titration sheet | RP1 | Permeabilized cells | Pc | »SUIT_RP1Pc | »SUIT_RP1Pc | 2016-07-01 |
Titration sheet | RP2 | Pc | »SUIT_RP2Pc | »SUIT_RP2Pc | 2016-07-01 | |
Tick boxes | Combined | Pc | »RP-Pc print | »RP-Pc print | ||
Titration sheet | RP1 | Isolated mitochondria, tissue homogenate, permeabilized fibers | Imt, Thom, Pfi | »SUIT_RP1mt | »SUIT_RP1mt | 2016-07-01 |
Titration sheet | RP2 | Imt, Thom, Pfi | »SUIT_RP2mt | »SUIT_RP2mt | 2016-07-01 | |
Tick boxes | Combined | Imt, Thom, Pfi | »RP print | »RP print |
Experimental details
Work in progress towards the next pre-publication version of »MiPNet21.06 SUIT reference protocol.pdf«
Saturating ADP concentrations
- Different concentrations of ADP are sufficient to obtain maximum flux for estimating OXPHOS capacity. The typical range for isolated mitochondria is 1 to 2.5 mM, for permeabilized cells 1 to 5 mM, for permeabilized muscle fibres 2.5 to 10 mM.
Pre-publication communication
- This communication is a pre-publication, inviting critical feedback and comments. All feedback will be carefully documented and evaluated in terms of justification of co-authorship. We intend to finally publish this topic in a peer-reviewed journal as an original article, with reference to the pre-publication history including reviewer's reports. - Gnaiger Erich 03:23, 18 January 2016 (CET)
Experiments in progress
- The SUIT reference protocol is presently applied in permeabilized HEK cells, mouse heart isolated mitochondria, liver homogenate, permeabilized skeletal muscle (mouse and human), and human PBMCs and platelets. - Gnaiger Erich 18:47, 19 January 2016 (CET)
- AT Innsbruck Gnaiger E and AT Innsbruck OROBOROS - MitoFit project.
- US NC Winston-Salem Molina AJA - UPBEAT project.
Further details
- » Introduction: Gnaiger 2014 MitoPathways
- » MitoPedia: SUIT - extended MitoPedia topic, May 2016
- Table of titrations » MiPNet09.12 O2k-Titrations
- » Definition: Substrate-uncoupler-inhibitor titration
- » Context: SUIT protocol library
- » Abbreviations: MitoPedia