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

MSCatalog No. MS1038

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Overview

Product name

Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit
See all Complex IV products (10) ...

Specificity

Species– Human. Rat and Mouse samples are not appropriate, other species are untested.

Precision

Intra-assay
Sample n Mean SD CV%
Overall 6 5.6%
Inter-assay
Sample n Mean SD CV%
Overall 6 10.9%

Tests

1 x 96 test

Sample type

Tissue Extracts, Cell Lysate

Assay type

Sandwich

Sensitivity

20 µg/ml

Recovery

Sample specific recovery
Sample type Average % Range %
Tissue Extracts 5 - 250
Cell Lysate 20 - 1000

Cross reactivity

Reacts with

Human

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.

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 applications

Sandwich ELISAmore details

Properties

Storage instructions

Please see Notes section

Components Identifier 1 x 96 tests
10X Blocking Buffer82098021 x 6ml
10X Cytochrome Oxidase Detector Antibody82095661 x 1ml
10X HRP Label82060381 x 1ml
20X Buffer82030131 x 20ml
Cytochrome Oxidase Microplate82095671 unit
Extraction Buffer82010931 x 15ml
TMB Development Solution82098031 x 12ml
  • Sandwich ELISA - Human Cytochrome Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538)Sandwich ELISA - Human Cytochrome Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538) image (enlarge)

  • Sandwich ELISA - Human Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538)Sandwich ELISA - Human Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538) image (enlarge)

  • Sandwich ELISA - Human Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538)Sandwich ELISA - Human Cytochrome-C Oxidase (Complex IV) Human Profiling ELISA Kit (ab124538) image (enlarge)

Alternative names

  • Cytochrome c oxidase
  • MS1038

Applications

Show applications key

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:

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

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

Example control sample curve of serially titrated HepG2 extract in the working range of the assay.

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

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

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.

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

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

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)

ab124538 has not yet been referenced specifically in any publications.

Publishing research using ab124538? Please let us know so that we can cite the reference in this datasheet

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