Key features and details
- Sensitivity: 4 µg/ml
- Range: 6 µg/ml - 200 µg/ml
- Sample type: Cell Lysate, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (qualitative)
- Reacts with: Human
Product nameHuman Cyt C Reductase ELISA Kit (Complex III) Profiling ELISA Kit
See all Complex III kits
Intra-assay Sample n Mean SD CV% Overall 8 4.7% Inter-assay Sample n Mean SD CV% Overall 6 19.7%
Sample typeTissue Extracts, Cell Lysate
Assay typeSandwich (qualitative)
Range6 µg/ml - 200 µg/ml
Assay durationMultiple steps standard assay
Species reactivityReacts with: Human
Does not react with: Mouse, Rat
Ubiquinol-cytochrome-c reductase (Complex III) is the third enzyme of the oxidative phosphorylation (OXPHOS) system within the mitochondrial inner membrane. Ubiquinol-cytochrome-c reductase is a large protein complex of approximately 250,000 MW made up of 11 different subunits. The enzyme forms a dimer in the mitochondrial inner membrane. The catalytic subunit cytochrome b is encoded on mitochondrial DNA (mtDNA). All other subunits are encoded by nuclear genomic DNA, made in the cytosol, and translocated into the organelle for assembly at the inner membrane. Complex III catalyses the pumping of protons across the inner membrane and transfer of electrons from ubiquinol to cytochrome c by a mechanism known as the Q cycle in which ubiquinol (Q) the electron carrier exists as a reduced, partially reduced or oxidized form.
ab124537 Cyt C Reductase (Complex III) human profiling kit is an in vitro enzyme-linked immunosorbent assay (ELISA) for the comparison of Complex III levels or profile in cell and tissue lysates. The assay employs a human Complex III specific antibody coated onto microplate well plate strips.
Samples are pipetted into the wells and Complex III present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti-Complex III detector antibody is added. After washing away unbound detector antibody, an HRP-conjugated secondary antibody specific for the detector antibody is pipetted into the wells. The wells are again washed, an HRP substrate solution (TMB) is added to the wells and color develops in proportion to the amount of Complex III bound. The developing blue color is measured at 600 nm.
Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.
Species– Human. Rat and Mouse samples are not appropriate, other species are untested.
Store all components at 4°C. This kit is stable for 6 months from receipt. Unused microplate strips should be returned to the pouch containing the desiccant and resealed.
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Blocking Buffer 1 x 6ml 10X Complex III Detector Antibody 1 x 1ml 10X HRP Label 1 x 1ml 20X Buffer 1 x 20ml Complex III Microplate 1 unit Extraction Buffer (ab260490) 1 x 15ml HRP Development Solution 1 x 12ml
RelevanceComplex II (succinate-ubiquinone oxidoreductase), one of the mitochondrial respiratory chain complexes, transfers electrons from succinate generated during the citric acid cycle to Complex III (ubiquinolcytochrome c oxidoreductase), via a mobile electron shuttle, ubiquinone. Complex III transfers electrons to Complex IV (cytochrome c oxidase) via another mobile electron shuttle, cytochrome c.
- coenzyme Q : cytochrome c — oxidoreductase
- cytochrome bc1 complex
Example control sample curve of serially titrated HepG2 extract in the working range of the assay.
Human HepG2 cells were cultured in chloramphenicol for 6 days to ensure a significant effect on mitochondrial DNA replication and mitochondrial protein translation, respectively. The antibiotic inhibited mitochondrial protein translation and assembly of Complexes I and IV but had no significant effect on Complex II, III or V.
Human HepG2 cells were cultured in NARTI Zalcitabine (ddC) for 6 days to ensure a significant effect on mitochondrial DNA replication and mitochondrial protein translation, respectively. The drug reduced mitochondrial DNA levels and hence mitochondrial protein expression. As a consequence the assembly of Complexes I, III and IV were severely affected. Note that loss of the two small mitochondrial DNA encoded subunits of Complex V (ATP synthase) does not affect overall assembly. Interestingly an increase in Complex II was induced as a consequence of I, III, IV loss possibly to up regulate mitochondrial citric acid cycle function.
ab124537 has been referenced in 6 publications.
- Huang GD et al. Moschamindole induces glioma cell apoptosis by blocking Mia40-dependent mitochondrial intermembrane space assembly and oxidative respiration. Phytother Res 35:3390-3405 (2021). PubMed: 33856743
- Cui P et al. STAT3 inhibition induced temozolomide-resistant glioblastoma apoptosis via triggering mitochondrial STAT3 translocation and respiratory chain dysfunction. Cell Signal 71:109598 (2020). PubMed: 32165236
- Jaworski S et al. Degradation of Mitochondria and Oxidative Stress as the Main Mechanism of Toxicity of Pristine Graphene on U87 Glioblastoma Cells and Tumors and HS-5 Cells. Int J Mol Sci 20:N/A (2019). PubMed: 30717385
- Khasawneh RR et al. Cross talk between 26S proteasome and mitochondria in human mesenchymal stem cells' ability to survive under hypoxia stress. J Physiol Sci 69:1005-1017 (2019). PubMed: 31679117
- Ye Q et al. Mitochondrial Effects of PGC-1alpha Silencing in MPP(+) Treated Human SH-SY5Y Neuroblastoma Cells. Front Mol Neurosci 10:164 (2017). ELISA ; Rat . PubMed: 28611589
- Cherry AB et al. Induced Pluripotent Stem Cells with a Pathological Mitochondrial DNA Deletion. Stem Cells N/A:N/A (2013). ELISA ; Human . PubMed: 23400930