Reference: C - SUIT-003
SUIT number: D013_ce1;ce1P;ce2Omy;ce3U;ce3Glc;ce3M;ce4Rot;ce5S;1Dig;1U;1c;2Ama;3AsTm;4Azd
SUIT-003 O2 ce-pce D013 is designed to study coupling control and plasma membrane permeability of living cells. Respiratory capacities are tested in a sequence of coupling states: ROUTINE, LEAK and Electron transfer pathway in the presence of glucose and malate. To study LEAK respiration, the phosphorylation system is 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. At Electron transfer pathway state, glucose is added to test the Crabtree effect on living cells. Malate is added to stimulate respiration of cells without intact plasma membrane (dead cells, dce). The viability index, VI, in the simple CCVP-Glc,M (SUIT-003 O2 ce-pce D013) may be underestimated if succinate uptake in the living cells proceeds through the dicarboxylate carrier of the plasma membrane. CCVP-Glc,M can be extended by the Complex IV module.
Compare SUIT protocols
MitoPedia methods: Respirometry