Fig 1: BN-PAGE for complex IV and OXPHOS Western blot in COX11 fibroblasts. Mitochondria isolated from Patient X1 and control fibroblasts analyzed by BN-PAGE immunoblotting for COX2, COX4, (a) and COX1 (b) suggest a decreased level of monomeric CIV more evident in mitochondria solubilized with Triton X-100, which separates the individual OXPHOS complexes (marked by *); ‡indicates previous COX1 signal and complex II (SDHA) was used as a loading control. (c) The levels of other OXPHOS subunits in isolated mitochondria were similar to controls (CI-NDUFA9, CIII-CORE1, and CV-ATP5A). (d) Representative SDS-PAGE and western blot analysis in whole cell lysates with an antibody cocktail targeting individual OXPHOS complex subunits showed consistently lower levels of CIV in patient X1; porin (VDAC1) was used as a loading control. BN, Blue native; CIV, complex IV; OXPHOS, oxidative phosphorylation; BN-PAGE, Blue native polyacrylamide gel electrophoresis; SDHA, succinate dehydrogenase complex subunit A; SDS, sodium dodecyl sulfate.
Fig 2: BN‐PAGE for complex IV and OXPHOS Western blot in COX11 fibroblasts. Mitochondria isolated from Patient X1 and control fibroblasts analyzed by BN‐PAGE immunoblotting for COX2, COX4, (a) and COX1 (b) suggest a decreased level of monomeric CIV more evident in mitochondria solubilized with Triton X‐100, which separates the individual OXPHOS complexes (marked by *); ‡indicates previous COX1 signal and complex II (SDHA) was used as a loading control. (c) The levels of other OXPHOS subunits in isolated mitochondria were similar to controls (CI‐NDUFA9, CIII‐CORE1, and CV‐ATP5A). (d) Representative SDS‐PAGE and western blot analysis in whole cell lysates with an antibody cocktail targeting individual OXPHOS complex subunits showed consistently lower levels of CIV in patient X1; porin (VDAC1) was used as a loading control. BN, Blue native; CIV, complex IV; OXPHOS, oxidative phosphorylation; BN‐ PAGE, Blue native polyacrylamide gel electrophoresis; SDHA, succinate dehydrogenase complex subunit A; SDS, sodium dodecyl sulfate.
Supplier Page from Abcam for Complex IV Human Enzyme Activity Dipstick Assay Kit