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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
- 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
- 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.
Compare SUIT protocols
References
MitoPedia concepts: MiP concept, SUIT protocol, Recommended
MitoPedia methods:
Respirometry
