MitoSciences (MS1038)

Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538)

Overview

  • Product nameCytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit
    See all Complex IV kits
  • Detection methodColorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 6 5.6%
    Overall 6 5.6%
  • Tests
    1 x 96 test
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay typeSandwich (qualitative)
  • Sensitivity
    20 µg/ml
  • Range
    5 µg/ml - 1000 µg/ml
  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Human

    Does not react with

    Mouse, 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.

  • Notes

    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 ELISAmore details
  • PlatformMicroplate

Properties

Applications

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.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

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.

Protocols

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) »

See all 3 Publications for this product

Product Wall

our current complex IV ELISA only works with human samples and does not cross react with rodents.

We analyzed Rat Liver Homogenate at concentrations from 0-250 microg/mL. We saw a low background but no dose response. It is possible with purified mitochondria, other tissue homogenates or with mouse samples there may be a significant signal.

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This is a profiling assay for relative comparison of treated and normal samples.

Cytochrome oxidase is a 13 subunit complex, the antibodies in this kit bind different subunits and hence measure the assembly of the complex rather than an indiv...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"