From Bioblast
Revision as of 12:50, 30 November 2023 by Grings Mateus (talk | contribs) (Created page with "{{MitoPedia |description=400px |info='''A''': protocol for simultaneous determination of O<sub>2</sub> flux and NADH autofluorescence in...")
Description
Reference: A: protocol for simultaneous determination of O2 flux and NADH autofluorescence in mitochondrial preparations (isolated mitochondria, tissue homogenate and permeabilized cells)- SUIT-006
SUIT number: D078_1mt;1PGM;2D;2anoxia;3Myx
O2k-Application: NADH
Communicated by Grings M, Cardoso Luiza HD (last update 2022-05-03)
Representative traces
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
mt | REN | mt | ||
1PGM | PGML(n) | N | CI | 1PGM
|
2D | PGMP | N | CI | 1PGM;2D
|
3Anox | N | CI | 1PGM;2D;3Anox | |
4Myx | N | CI | 1PGM;2D;3Anox;4Myx
| |
5Reox | ROX | 1PGM;2D;3Anox;4Myx;5Reox |
- 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
Compare SUIT protocols
Chemicals and syringes
Step | Chemical(s) and link(s) | Comments |
---|---|---|
1PGM | Pyruvate (P), Glutamate (G), and Malate (M) | |
2D | ADP (D) | |
3Anox | The O2 concentration in the O2k-chamber can be decreased by N2 or H2 injection to reach faster anoxia, see: Setting the oxygen concentration. | |
4Myx | Myxothiazol | We do not recommend the use of any other inhibitor of complex III, like Antimycin A (Ama), due to the chemical background effect on fluorescence. |
5Reox | Reoxygenation can be performed by opening the chamber, see: Open chamber. |
- Suggested stock concentrations are shown in the specific DL-Protocol.