Difference between revisions of "SUIT-009 O2 ce-pce D016"
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|mitopedia method=Respirometry | |||
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::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(P) | ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS(P) | ||
::: '''[[SUIT protocol pattern]]:''' orthogonal 1S;2D;3P;4Rot;5Ama | ::: '''[[SUIT protocol pattern]]:''' orthogonal 1S;2D;3P;4Rot;5Ama | ||
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::: +This protocol can be extended with the Complex IV module. | ::: +This protocol can be extended with the Complex IV module. | ||
:::- In some cell lines e.g. HEK293T, P does not have any effect because of the downregulation of the N-linked pathway. | :::- In some cell lines e.g. HEK293T, P does not have any effect because of the downregulation of the N-linked pathway. | ||
Revision as of 15:25, 24 January 2019
Description
Abbreviation: NS(P)
Reference: A: SUIT-009 short protocol for mt-respiration in pce.
SUIT number: D016_ce1;1Dig;1S;2D;3P;4Rot;5Ama
MitoPedia concepts:
SUIT protocol,
SUIT B,
Find
MitoPedia methods:
Respirometry
- SUIT-category: NS(P)
- SUIT protocol pattern: orthogonal 1S;2D;3P;4Rot;5Ama
SUIT 9 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. Succinate (S) supports the reverse electron transfer (RET) initiated H2O2 production in the LEAK state. Addition of ADP initiates S-OXPHOS capacity. NS pathway control is induced by addition of succinate and NADH-linked substrates such as Pyruvate (P). For O2-Application, choose SUIT-009 O2 mt D015; for O2 and H2O2 measurements (AmR), choose: D021_1S;2D;3P;4Rot;5Ama.
Communicated by Iglesias-Gonzalez J, Komlodi T and Gnaiger E (last update 2019-01-23)
SUIT protocols
References
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
1Dig | Dig | ce1;1Dig
|
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
1S | SL(n) | S | CII | 1S
|
2D | SP | S | CII | 1S;2D
|
2c | ScP | S | CII | 1S;2D;2c
|
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 pce D019; for O2 and H2O2 measurements (AmR), choose: SUIT-009 AmR pce D019.
- 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.
- - In some cell lines e.g. HEK293T, P does not have any effect because of the downregulation of the N-linked pathway.