Table 1.
Substrates, uncoupler, and inhibitors used in respirometry protocol to induce different respiratory states [20,21].
| Substrates | Function | Concentration |
|---|---|---|
| Digitonin (Dig) | Plasma membrane permeabilization | 7.5 µg∙mL−1 (NALCL) 27.5 µg∙mL−1 (ALCL) |
| Pyruvate (P) | NADH-generating substrate (Substrates for CI) | 5 mM |
| Glutamate (G) | 10 mM | |
| Malate (M) | 2 mM | |
| Succinate (S) | CII substrate | 10 mM |
| Ascorbate (As) | Maintains TMPD in a reduced state | 2 mM |
| N,N,N′,N′-tetramethyl-p-phenylenediamine dihydrochloride (TMPD) | Substrate for reducing cytochrome c (Substrate for CIV) | 0.5 mM |
| Cytochrome c (c) | Mitochondrial outer membrane permeability test | 10 µM |
| ADP (D) | Substrate of ANT, F1F0-ATPase (Substrate for CV) | 2.5 mM |
| Carbonyl cyanide m-chlorophenyl hydrazone, CCCP (U) | Uncoupler, protonophore | 0.5 µM steps |
| Rotenone (Rot) | CI inhibitor | 0.5 µM |
| Malonate (Mna) | CII inhibitor | 5 mM |
| Antimycin A (Ama) | CIII inhibitor | 2.5 µM |
| Sodium azide (Azd) | CIV inhibitor | 100 mM |
| Oligomycin (Omy) | ATP synthase inhibitor | 2.5 µM |
| Safranin (Saf) | Fluorophore, dye for measuring mitochondrial membrane potential | 2 µM |