Talk:SUIT-031

high-resolution terminology - matching measurements at high-resolution

Talk:SUIT-031

Description

Abbreviation: PM+S+Rot_OXPHOS

Reference: A: simultaneous determination of O₂ flux and the redox state of the Q-pool

SUIT-category: NS

SUIT protocol pattern: orthogonal 1PM;2D;3S;4Rot;5U;-

This is a protocol to investigate simultanouesly oxygen flux and the redox state of the Q-pool in the N- and NS-pathway control state on isolated mitochondria and permeabilized cells.

Communicated by  Komlodi T (names to be added) (last update 2021-02-01)

Specific SUIT protocols

SUIT-031 Q mt D072

File:1PM;2D;2S;4Rot;5U;6Ama.png

• SUIT-031 Q mt D072 simultaneous determination of O₂ flux and the redox ratio of the Q-pool in isolated mitochondria.

SUIT-031 Q ce-pce D074

File:Ce1;1Dig;1PM;2D;2S;4Rot;5U;6Ama.png

• SUIT-031 Q ce-pce D074 simultaneous determination of O₂ flux and the redox ratio of the Q-pool in permeabilized cells.

SUIT-031 O2 mt D075

Step	Chemical(s) and link(s)	Comments
1PM	Pyruvate (P) and Malate (M)
2D	ADP (D)
3S	Succinate (S)
4Rot	Rotenone (Rot)
5U	Carbonyl cyanide m-chlorophenyl hydrazone, CCCP (U)	Can be substituted for other uncoupler.
6Ama	Antimycin A (Ama)

Strengths and limitations

• NS-OXPHOS capacity provides a physiologically relevant estimate of maximum mitochondrial respiratory capacity.
• A succinate concentration of >10 mM may be required for saturating SE capacity.

+ You obtain information in a single protocol about the NS-, S and N- pathway control state.

- CIV activity cannot be determined and Cytochrome c test cannot be performed together with the fluorescence dyes.

- Careful washing is required after the experiment to avoid carry-over of the inhibitors and uncoupler.

References

MitoPedia concepts: MiP concept, SUIT protocol, Recommended 

MitoPedia methods: Respirometry