SUIT-003 O2 ce D009

high-resolution terminology - matching measurements at high-resolution

SUIT-003 O2 ce D009

Description

Abbreviation: RLU

Reference: A Huetter 2004 Biochem J

MitoPedia concepts: SUIT protocol, SUIT A 

SUIT-category: RLU-

SUIT protocol pattern: linear coupling control protocol (CCP)

References

DatLab-Analysis template: CCP02.xlsx

• The CCP diagram may show the respiratory states directly (left) as an alternative to presenting the titration steps.
• Rot and Ama

In this CCP, 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 (4Ama).

CCP02-Rot,Ama

• CCP02_R,1Omy,2U,3Rot-4Ama
• Rot and Ama

In this CCP, ROX is measured after stepwise titration of rotenone (3Rot) and antimycin A (4Ama). In many intact cells ROX measured after titration of rotenone (3Rot) is not or only slightly further inhibited by subsequent titration of animycin A (4Ama).

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: ROUTINE (R; routine state in cell-culture medium); LEAK (L; inhibition of ATPsynthase by 1 μg/ml oligomycin); ETS (E; maximal stimulation by uncoupling of oxidative phosphorylation in four subsequent titrations of the uncoupler, U, FCCP (2.5–4 μM final concentration); ROX (rotenone, Rot, and antimycin A; residual oxygen consumption). Modified after Huetter_2004_Biochem J.

• 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«