Abbreviation: CCP-ce Snv - control
SUIT number: D062_ce1;ce1DMSO;(ce2Omy);ce3U;ce4Rot;ce5Ama
SUIT-003 O2 ce D062 protocol is the control of the SUIT-003 O2 ce D050 adding the carrier DMSO in ROUTINE respiration instead of the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv). Moreover,the SUIT-003 O2 ce D062 protocol allows to evaluate the coupling control of living cells. Respiratory capacities are tested in a sequence of coupling states: ROUTINE and Electron transfer pathway. Optionally, to study LEAK respiration, the phosphorylation system might be inhibited by oligomycin. The final concentration of oligomycin has to be carefully optimized for various cell types, to minimize the inhibitory effect on the electron transfer system which would lead to an underestimation of ET capacity.
Communicated by Doerrier C (last update 2019-06-26)
Steps and respiratory states
|Step||State||Pathway||Q-junction||Comment - Events (E) and Marks (M)
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- Pathway control
Strengths and limitations
- + This is a cell coupling-control protocol that provides information on ROUTINE and Electron transfer pathway in the presence of (MitoKit-CII/Succinate-nv.
- - This protocol is provided without oligomycin addition. To also include LEAK respiration in the protocol, an event for the addition of oligomycin and a related mark can be added by editing the DLP file, creating a DLPU. However, the addition of oligomycin has to be carefully tested to avoid inhibitory effects in the evaluation of the ET capacity.
- - Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.
Compare SUIT protocols
- SUIT-003 O2 ce D050 the plasma membrane-permeable succinate (MitoKit-CII/Succinate-nv) is added in ROUTINE.
MitoPedia methods: Respirometry