Difference between revisions of "SUIT-033 NADH mt D081"
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|info='''A''': protocol for simultaneous determination of O<sub>2</sub> flux and NADH autofluorescence in mitochondrial preparations (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-006]]''' | |info='''A''': protocol for simultaneous determination of O<sub>2</sub> flux and NADH autofluorescence in mitochondrial preparations (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-006]]''' | ||
|application=NADH | |application=NADH | ||
|SUIT number= | |SUIT number=D081_mt;1D.1;2PGM;3D2.5;4Anox;5Myx;6Reox | ||
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Revision as of 18:09, 13 December 2023
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: D081_mt;1D.1;2PGM;3D2.5;4Anox;5Myx;6Reox
O2k-Application: NADH
Communicated by Grings M, Cardoso Luiza HD (last update 2023-11-30)
Representative traces
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
mt | REN | mt | ||
1D.1 | mt;1D.1
| |||
2PGM | PGM | N | CI | mt;1D.1;2PGM
|
3D2.5 | PGMP | N | CI | mt;1D.1;2PGM;3D2.5
|
4Anox | N | CI | mt;1D.1;2PGM;3D2.5;4Anox | |
5Myx | N | CI | mt;1D.1;2PGM;3D2.5;4Anox;5Myx
| |
6Reox | ROX | mt;1D.1;2PGM;3D2.5;4Anox;5Myx;6Reox |
- 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 |
---|---|---|
1D.1 | ADP (D) | |
2PGM | Pyruvate (P), Glutamate (G), and Malate (M) | |
3D2.5 | ADP (D) | |
4Anox | The O2 concentration in the O2k-chamber can be decreased by N2 or H2 injection to reach faster anoxia, see: Setting the oxygen concentration. | |
5Myx | 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. |
6Reox | Reoxygenation can be performed by opening the chamber, see: Open chamber. |
- Suggested stock concentrations are shown in the specific DL-Protocol.