Difference between revisions of "SUIT-003 O2 ce D009"
From Bioblast
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::: '''[[SUIT protocol pattern]]:''' linear cell (ce) [[coupling control protocol]] (ceCCP) | ::: '''[[SUIT protocol pattern]]:''' linear cell (ce) [[coupling control protocol]] (ceCCP) | ||
== SUIT O2 DLP for SUIT 3 (DatLab 7.3) == | |||
''Updating this page (2018-06-13) for DatLab 7.3 DLP files'' | |||
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[[File:1ce;2ceOmy;3ceU;4ceRot;5ceAma.jpg|right|200px]] | [[File:1ce;2ceOmy;3ceU;4ceRot;5ceAma.jpg|right|200px]] | ||
== | == ce1;ce2Omy;ce3U;ce4Rot;ce5Ama == | ||
:::* | :::* ce1;ce2Omy;ce3U;ce4Rot;ce5Ama | ||
:::: In this ceCCP, ROX is measured after stepwise titration of [[rotenone]] (ce4Rot) and [[antimycin A]] (ce5Ama). In many intact cells ROX measured after titration of rotenone (ce4Rot) is not or only slightly further inhibited by subsequent titration of animycin A (ce5Ama). | :::: In this ceCCP, ROX is measured after stepwise titration of [[rotenone]] (ce4Rot) and [[antimycin A]] (ce5Ama). In many intact cells ROX measured after titration of rotenone (ce4Rot) is not or only slightly further inhibited by subsequent titration of animycin A (ce5Ama). |
Revision as of 15:19, 13 June 2018
Description
Reference: A Huetter 2004 Biochem J
SUIT number: 3
O2k-Application: O2
MitoPedia concepts:
SUIT protocol,
SUIT A
- SUIT protocol pattern: linear cell (ce) coupling control protocol (ceCCP)
SUIT O2 DLP for SUIT 3 (DatLab 7.3)
Updating this page (2018-06-13) for DatLab 7.3 DLP files
References
R;L;E format_ROX
- The ceCCP diagram may show the respiratory states directly (left) as an alternative to presenting the titration steps.
- Rot and Ama
- In this ceCCP, ROX is measured after titration of rotenone, antimycin A, or both in a single step. Validity of using rotenone only for inducing ROX in intact cells is evaluated by stepwise measurement of ROX after titration of rotenone (3Rot) followed by titration of animycin A (4ceAma).
ce1;ce2Omy;ce3U;ce4Rot;ce5Ama
- ce1;ce2Omy;ce3U;ce4Rot;ce5Ama
- In this ceCCP, ROX is measured after stepwise titration of rotenone (ce4Rot) and antimycin A (ce5Ama). In many intact cells ROX measured after titration of rotenone (ce4Rot) is not or only slightly further inhibited by subsequent titration of animycin A (ce5Ama).
- Respiration in senescent human primary fibroblasts (0.2×106 cells/ml). Arrows show steps in the titration regime of the coupling control protocol, inducing the following respiratory states.
- 1ce: ROUTINE (R; routine state in cell-culture medium) in intact cells (ce);
- 2ceOmy: LEAK (L; inhibition of ATPsynthase by 1 μg/ml oligomycin);
- 3ceU: ET-pathway (E; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5–4 μM final concentration);
- 4ceRot: ROX (rotenone, Rot; residual oxygen consumption).
- 5ceAma: ROX (antimycin A, Ama; residual oxygen consumption). Modified after Huetter_2004_Biochem J.
- Respiration in senescent human primary fibroblasts (0.2×106 cells/ml). Arrows show steps in the titration regime of the coupling control protocol, inducing the following respiratory states.
- Hütter E, Renner K, Pfister G, Stöckl P, Jansen-Dürr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts. Biochem J 380:919-28. - »Bioblast link«