Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538)
- Product nameCytochrome-C Oxidase (Complex IV) Human Profiling ELISA KitSee all Complex IV kits ...
- Detection methodColorimetric
Intra-assay Sample n Mean SD CV% Overall 6 5.6%
- Tests1 x 96 test
- Sample typeTissue Extracts, Cell Lysate
- Assay typeSandwich (qualitative)
- Sensitivity20 µg/ml
- Range5 µg/ml - 1000 µg/ml
- Assay durationMultiple steps standard assay
- Species reactivityReacts with: Human
Does not react withMouse, Rat
- Product overview
Cytochrome-c oxidase (Complex IV) is the forth enzyme of the oxidative phosphorylation (OXPHOS) system within the mitochondrial inner membrane. Cytochrome-c oxidase is a large protein complex of approximately 200,000 MW made up of 13 different subunits. Three catalytic subunits I,II, and III are 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 IV catalyses the pumping of protons across the inner membrane and the transfer of electrons to the terminal acceptor oxygen (O2) making water. Several cytochrome-c oxidase subunits have isoforms present in different tissues which subtly regulate enzymatic activity. Isoform expression or switching is likely in response to intracellular oxygen concentration.
ab124538 Cytochrome-c oxidase (Complex IV) Human Profiling kit is an in vitro enzyme-linked immunosorbent assay (ELISA) for the comparison of cytochrome-c oxidase levels or profile in cell and tissue lysates. The assay employs a human cytochrome-c oxidase specific capture antibody coated onto microplate well strips.
Samples are pipetted into the wells and cytochrome-c oxidase present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti- cytochrome-c oxidase 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 cytochrome-c oxidase 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.
- Tested applicationsSandwich ELISA more details
- Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Blocking Buffer 1 x 6ml 10X Cytochrome Oxidase Detector Antibody 1 x 1ml 10X HRP Label 1 x 1ml 20X Buffer 1 x 20ml Cytochrome Oxidase Microplate 1 unit Extraction Buffer 1 x 15ml TMB Development Solution 1 x 12ml
- Research Areas
- Cytochrome c oxidase
- Cytochrome oxidase
- Entrez Gene: 4512 Human
- Entrez Gene: 4513 Human
- Entrez Gene: 4514 Human
- Entrez Gene: 1345 Human
- Entrez Gene: 1351 Human
- Entrez Gene: 1329 Human
- Entrez Gene: 1349 Human
- Entrez Gene: 1337 Human
- Entrez Gene: 1327 Human
- Entrez Gene: 1340 Human
- Entrez Gene: 1350 Human
- Entrez Gene: 9377 Human
- Entrez Gene: 1346 Human
- SwissProt: P00395 Human
- SwissProt: P00403 Human
- SwissProt: P00414 Human
- SwissProt: P13073 Human
- SwissProt: P12074 Human
- SwissProt: P15954 Human
- SwissProt: P24311 Human
- SwissProt: P10176 Human
- SwissProt: P14854 Human
- SwissProt: P09669 Human
- SwissProt: P24310 Human
- SwissProt: P20674 Human
- SwissProt: P10606 Human
Our Abpromise guarantee covers the use of ab124538 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit images
Example control sample curve of serially titrated HepG2 extract in the working range of the assay.
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. 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.
Human HepG2 cells were cultured in chloramphenicol for 6 days to ensure a significant effect on mitochondrial DNA replication and mitochondrial protein translation. The antibiotic inhibited mitochondrial protein translation and assembly of Complexes I and IV but had no significant effect on Complex II, III or V.
References for Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538)
This product has been referenced in:
- Gómez-Durán A et al. Oxidative phosphorylation differences between mitochondrial DNA haplogroups modify the risk of Leber's hereditary optic neuropathy. Biochim Biophys Acta 1822:1216-22 (2012). Read more (PubMed: 22561905) »
- Schmidt H et al. 2-D DIGE analyses of enriched secretory lysosomes reveal heterogeneous profiles of functionally relevant proteins in leukemic and activated human NK cells. Proteomics 8:2911-25 (2008). Read more (PubMed: 18655029) »