MiPNet21.06 SUIT RP

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 

Work in progress

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 names is very restricted. The simplest solution for short mark names is the use of a numerical sequence, with the event name added for information.

• RP1: 1PM 2D 3c 4U 5Oct 6G 7S 8Rot 9Gp 10Ama 11Tm 12Azd

~ Gnaiger Erich 16:18, 22 January 2016 (CET)

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

» MitoPedia: Respiratory substrate-coupling states - extended MitoPedia topic, Jan 2016

• Table of titrations » MiPNet09.12 O2k-Titrations
• Definition » Substrate-uncoupler-inhibitor titration
• Context » SUIT protocol library
• Abbreviations » MitoPedia