Difference between revisions of "MiPNet21.06 SUIT RP"
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This communication is a pre-publication, inviting critical feedback and comments. All feedback will be carefully documented and evaluated in terms of justification of [[Gentle_Science#The_Vancouver_Protocol.2C_ICMJE.2C_and_Gentle_Science |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. - [[User:Gnaiger Erich|Gnaiger Erich]] 03:23, 18 January 2016 (CET) | This communication is a pre-publication, inviting critical feedback and comments. All feedback will be carefully documented and evaluated in terms of justification of [[Gentle_Science#The_Vancouver_Protocol.2C_ICMJE.2C_and_Gentle_Science |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. - [[User:Gnaiger Erich|Gnaiger Erich]] 03:23, 18 January 2016 (CET) | ||
=== Experiments in progress === | |||
The '''SUIT reference assay''' is presently applied in permeabilized HEK cells, mouse heart isolated mitochondria, liver homogenate, permeabilized skeletal muscle (mouse and human), and human PBMCs and platelets. - [[User:Gnaiger Erich|Gnaiger Erich]] 18:47, 19 January 2016 (CET) | The '''SUIT reference assay''' is presently applied in permeabilized HEK cells, mouse heart isolated mitochondria, liver homogenate, permeabilized skeletal muscle (mouse and human), and human PBMCs and platelets. - [[User:Gnaiger Erich|Gnaiger Erich]] 18:47, 19 January 2016 (CET) | ||
* [[AT Innsbruck Gnaiger E]] and [[AT Innsbruck OROBOROS]] - [[O2k-MitoFit |MitoFit]] project. | * [[AT Innsbruck Gnaiger E]] and [[AT Innsbruck OROBOROS]] - [[O2k-MitoFit |MitoFit]] project. |
Revision as of 19:57, 19 January 2016
SUIT reference assay for OXPHOS analysis by high-resolution respirometry. - »Bioblast pdf« |
» Versions
OROBOROS (2016-01-19) Mitochondr Physiol Network
Abstract: Doerrier C, Sumbalova Z, Lamberti G, Krumschnabel G, Gnaiger (2016) SUIT reference assay for OXPHOS analysis by high-resolution respirometry. Mitochondr Physiol Network 21.06(pre03):1-10.
- » 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, Protocol"Protocol" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property.
O2k-SOP
Laboratory sheets for SUIT protocols RP1 and RP2
Type | SUIT protocol | Sample | Abbr | pdf file | docx file |
---|---|---|---|---|---|
Titration sheet | RP1 | Permeabilized fibres | Pfi | »SUIT_RP1-Pfi | »SUIT_RP1-Pfi |
Titration sheet | RP2 | Pfi | »SUIT_RP2-Pfi | »SUIT_RP2-Pfi | |
Tick boxes | Combined | Pfi | »RP print-Pfi | »RP print-Pfi | |
Titration sheet | RP1 | Permeabilized cells | Pc | »SUIT_RP1 | »SUIT_RP1 |
Titration sheet | RP2 | Pc | »SUIT_RP2 | »SUIT_RP2 | |
Tick boxes | Combined | Pc | »RP-Pc print | »RP-Pc print | |
Titration sheet | RP1 | Isolated mitochondria, tissue homogenate | Imt, Thom | »SUIT_RP1 | »SUIT_RP1 |
Titration sheet | RP2 | Imt, Thom | »SUIT_RP2 | »SUIT_RP2 | |
Tick boxes | Combined | Imt, Thom | »RP print | »RP print |
Experimental details
Work in progress towards the next pre-publication version of »MiPNet21.06 SUIT reference assay.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 assay 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
- Table of titrations » MiPNet09.12 O2k-Titrations
- Definition » Substrate-uncoupler-inhibitor titration
- Context » SUIT protocol library
- Abbreviations » MitoPedia