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
Originially this 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. It is used as a control experiment forSUIT-009 AmR mt D021 without using the Amplex UltraRed assay in order to detect the effect of the fluorescence dye on the O2 flux. We do not recommend this protocol for determination of the O2 flux in the absence of Rotenone. For O2-Application with permeabilized cells, choose SUIT-009 O2 pce D016; for O2 and H2O2 measurements (AmR) wiht isoalted mitochondria and tissue homogenate, choose: SUIT-009 AmR mt D021.
Communicated by Iglesias-Gonzalez J, Komlodi T and Gnaiger E (last update 2019-01-22)
DLP applications
- SUIT-009 AmR mt D021 simultaneous determination of O2 and H2O2 flux on isolated mitochondria and tissue homogenate.
- SUIT-009 O2 mt D015 determination of O2 flux on isolated mitochondria and tissue homogenate as a control for SUIT-009 AmR mt D021.
- SUIT-009 AmR pce D019 simultaneous determination of O2 and H2O2 flux on permeabilized cells.
- SUIT-009 O2 pce D016 determination of O2 flux on permeabilized cells as a control for SUIT-009 AmR pce D019.
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 |
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- Coupling control
- Pathway control
- Main fuel substrates
- » Glutamate, G
- » Glycerophosphate, Gp
- » Malate, M
- » Octanoylcarnitine, Oct
- » Pyruvate, P
- » 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 B, Find
MitoPedia methods:
Respirometry