SUIT-006 NADH mt D084: Difference between revisions
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|description=[[File:1PGM;2D;3U;3anoxia;4Myx.png|400px]] | |description=[[File:1PGM;2D;3U;3anoxia;4Myx.png|400px]] | ||
|info='''A''':ย 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''':ย 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]]''' | ||
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|SUIT number=D084_1PGM;2D;3U;3anoxia;4Myx | |SUIT number=D084_1PGM;2D;3U;3anoxia;4Myx | ||
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ย | {{MitoPedia concepts}} | ||
ย | {{MitoPedia methods}} | ||
{{MitoPedia O2k and high-resolution respirometry}} | |||
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ย Communicated byย [[Doerrier C]] (last update 2022-05-03) ย | ย Communicated byย [[Doerrier C]] (last update 2022-05-03) ย | ||
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== Chemicals and syringes == | == Chemicals and syringes == | ||
{{Template:Chemical SUIT-006 NADH mt D084}} | |||
::: Suggested stock concentrations are shown in the specific DL-Protocol. | ::: Suggested stock concentrations are shown in the specific DL-Protocol. | ||
Revision as of 12:39, 30 November 2023
Description
File:1PGM;2D;3U;3anoxia;4Myx.png
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: D084_1PGM;2D;3U;3anoxia;4Myx
O2k-Application: NADH
Communicated by Doerrier C (last update 2022-05-03)
Representative traces
File:SUIT-006 NADH mt D084 O2.png File:SUIT-006 NADH mt D084.png
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
|
(3Omy) | PGML(Omy) | N | CI | 1PGM;2D;(3Omy)
|
4U | PGME | N | CI | 1PGM;2D;(3Omy);4U
|
5Anox | N | CI | 1PGM;2D;(3Omy);4U;5Anox | |
6Myx | N | CI | 1PGM;2D;(3Omy);4U;5Anox;6Myx
| |
7Reox | ROX | 1PGM;2D;(3Omy);4U;5Anox;6Myx;7Reox |
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- Coupling control
- Pathway control
- ยป Electron transfer pathway
- ยป Fatty acid oxidation pathway control state, F
- ยป NADH electron transfer-pathway state, N
- ยป Succinate pathway control state, S
- ยป NS-pathway control state, NS
- ยป Glycerophosphate pathway control state, Gp
- ยป Complex IV single step, CIV
- ยป Anaplerotic pathway control state
- Pathway control
- Main fuel substrates
- ยป Glutamate, G
- ยป Glycerophosphate, Gp
- ยป Malate, M
- ยป Octanoylcarnitine, Oct
- ยป Pyruvate, P
- ยป 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) | |
(3Omy) | Oligomycin (Omy) | This step can be skipped. |
4U | SF6847 | We do not recommend the use of any other uncoupler, like Carbonyl cyanide m-chlorophenyl hydrazone, CCCP (U), due to the chemical background effect on fluorescence. |
5Anox | The O2 concentration in the O2k-chamber can be decreased by N2 or H2 injection to reach faster anoxia, see: Setting the oxygen concentration. | |
6Myx | 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. |
7Reox | Reoxygenation can be performed by opening the chamber, see: Open chamber. |
- Suggested stock concentrations are shown in the specific DL-Protocol.