From Bioblast
Description
Abbreviation: NS(PGM)
Reference: A: SUIT-020 short protocol for simultaneous determination of O2 flux and mitochondrial membrane potential in mitochondrial preparations (isolated mitochondria and tissue homogenate).
SUIT number: D033_1PM;2D;3G;4S;5Rot;6Omy;7U;8Ama
O2k-Application: AmR
- SUIT-category: NS(PGM)
- SUIT protocol pattern: orthogonal 1PM;2D;3G;4S;5Rot;6Omy;7U;8Ama
Communicated by Komlodi T (last update 2019-01-28)
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1PM | PML(n) | N | CI | 1PM
|
2D | PMP | N | CI | 1PM;2D
|
3G | PGMP | N | CI | 1PM;2D;3G
|
4S | PGMSP | NS | CI&II | 1PM;2D;3G;4S
|
5Rot | SP | S | CII | 1PM;2D;3G;4S;5Rot
|
6Omy | SL(Omy) | 1PM;2D;3G;4S;5Rot;6Omy
| ||
7U* | SE | S | CII | 1PM;2D;3G;4S;5Rot;6Omy;7S
|
8Ama | ROX | 1PM;2D;3G;4S;5Rot;6Omy;7S;8Ama
|
- Bioblast links: SUIT protocols - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>
- 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
- + Information in one protocol about the NS, S and N-pathway control state.
- - The fluorescence dye (such as Safranin, TMRM etc) can inhibit most porbably the NADH-linked respiration, therefore, the protocol has to be run without using the fluorescence dye. The following protocols can be used: SUIT-020 O2 mt D032, SUIT-020 O2 mt D035.
- - CIV activity and Cytochrome c test cannot be performed together with the fluorescence.
- - Omy concentration has to be determined. Higher concentration of Omy than required can inhibit the ET state.
Compare SUIT protocols
- SUIT-004;SUIT-004 O2 pfi D010: it provides an information about the NS(PM) pathway in the ET state withou the contribution of G.
- SUIT-008;SUIT-008 O2 pfi D014 and SUIT-008 O2 pce D025: it provides information about the NS(PGM) pathway in the OXPHOS and ET-states, but without the addtion of Omy.
- SUIT-011:it provides information about the NS(GM) pathway in the OXPHOS and ET-states, but without the addtion of Omy.
- SUIT-012:it provides information about the N(PGM) pathway in the OXPHOS and ET-states wihtout the contribution of S-pathway and without the addtion of Omy.
- SUIT-014: it is similar protocol to SUIT-020 O2 mt D035, but with the addition of P in the OXPHOS state before S. Omy is not added in the protocol.
References
Year | Reference | Organism | Tissue;cell | |
---|---|---|---|---|
MiPNet20.13 Safranin mt-membranepotential | 2019-06-24 | O2k-FluoRespirometry: HRR and simultaneous determination of mt-membrane potential with safranin or TMRM. | Mouse | Nervous system |
Krumschnabel 2014 Methods Enzymol | 2014 | Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2014) Use of safranin for the assessment of mitochondrial membrane potential by high-resolution respirometry and fluorometry. Methods Enzymol 542:163-81. https://doi.org/10.1016/B978-0-12-416618-9.00009-1 | Mouse | Nervous system |
MitoPedia concepts:
SUIT protocol,
SUIT A,
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MitoPedia methods:
Fluorometry