Description
Abbreviation: NS(P)
Reference: A: SUIT-009 short protocol for respiration in mitochondrial preparations (isolated mitochondria and tissue homogenate).
SUIT number: D015_1S;2D;3P;4Rot;5Ama
- SUIT-category: NS(P)
- SUIT protocol pattern: orthogonal 1S;2D;3P;4Rot
SUIT 9 is a short protocol for simultaneous determination of O2 flux and the rate of H2O2 production (H2O2 flux) on isolated mitochondria,tissue homogenate (except of liver) and permeabilized cells. Succinate (S) supports the reverse electron transfer (RET) initiated H2O2 production in the LEAK state. Addition of ADP initiates S-OXPHOS capacity. NS pathway control is induced by addition of succinate and NADH-linked substrates such as Pyruvate (P). For O2-Application, choose SUIT-009 O2 mt D015; for O2 and H2O2 measurements (AmR), choose: D021_1S;2D;3P;4Rot;5Ama.
Communicated by Iglesias-Gonzalez J, Komlodi T and Gnaiger E (last update 2019-01-22)
DLP applications
References
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1S | SL(n) | S | CII | 1S
|
2D | SP | S | CII | 1S;2D
|
3P | NSP | NS | CI&II | 1S;2D;3P
|
4Rot | SP | S | CII | 1S;2D;3P;4Rot
|
5Ama | ROX | 1S;2D;3P;4Rot;5Ama
|
Step | Respiratory state | Pathway control | ET-Complex | Comment |
---|---|---|---|---|
## AsTm | AsTmE | CIV | CIV | |
## Azd | CHB |
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- Coupling control
- Pathway control
- Β» Electron transfer pathway
- Β» Fatty acid oxidation pathway control state, F
- Β» NADH electron transfer-pathway state, N
- Β» Succinate pathway control state, S
- Β» NS-pathway control state, NS
- Β» Glycerophosphate pathway control state, Gp
- Β» Complex IV single step, CIV
- Β» Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- Β» Glutamate, G
- Β» Glycerophosphate, Gp
- Β» Malate, M
- Β» Octanoylcarnitine, Oct
- Β» Pyruvate, P
- Β» Succinate, S
- Main fuel substrates
- Glossary
Strengths and limitations
- For H2O2-Application, choose SUIT-009 AmR mt D021; for O2 and H2O2 measurements (AmR), choose: SUIT-009 AmR mt D021.
- Cytochrome c test can be performed in the S-OXPHOS state.
- +Reasonable duration of the experiment.
- +This protocol can be extended with the Complex IV module.
Compare SUIT protocols
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry