From Bioblast
Description
Abbreviation: ceGlc
Reference: A Coupling control protocol and Crabtree effect test -SUIT-003
SUIT number: D037_ce1;ce2Glc;ce3U
O2k-Application: O2
- MitoPedia: SUIT
- SUIT-category: ceGlc
- SUIT protocol pattern: complex cell coupling control protocol ce1;ce2Glc;ce3U
SUIT-003 O2 ce D037 has been designed to study the coupling control state and respiration regulation by glucose (Crabtree effect) of intact cells on ROUTINE. The different capacities are tested in the sequence OXPHOS and ET. SUIT-003 O2 ce D037 follows a modular approach and can be easily used in combination with the assessment of complex IV capacity and respirometric cell viability test.
Steps and respiratory states
Step | State | Pathway | Q-junction | Comment - Events (E) and Marks (M) |
---|---|---|---|---|
ce1 | ROUTINE | ce1
| ||
ce1Glc | ROUTINE | ce1;ce1Glc
| ||
(ce2Omy) | L(Omy) | ce1;ce1Glc;(ce2Omy)
| ||
ce3U | E | ce1;ce1Glc;(ce2Omy);ce3U
| ||
ce4Ama | ROX | ce1;ce1Glc;(ce2Omy);ce3U;ce4Ama
|
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- 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
- The SUIT-003 O2 ce D037 is a cell coupling control protocol. The total cell count (Nce) is the sum of viable cells (Nvce) and dead cells (Ndce).
- This protocol can be extended with the cell viability test module (See CCV SUIT-003 O2 ce-pce D020).
- CIV activity can also be measured after the cell viability test module.
- + Reasonable duration of the experiment.
- - Cell culture media with glucose (e.g. DMEM: 25 Β΅M Glucose) could hinder the cell viability test.
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
Compare SUIT protocols
References
MitoPedia concepts: SUIT protocol, SUIT A, Find
MitoPedia methods:
Respirometry