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

Overview

• Product nameCytochrome-C Oxidase (Complex IV) Human Profiling ELISA KitSee all Complex IV kits ...
• Precision

  Intra-assay

  Sample	n	Mean	SD	CV%
  Overall	6			5.6%
• Tests
  1 x 96 test
• Sample type
  Tissue Extracts, Cell Lysate
• Assay typeSandwich
• Sensitivity
  20 µg/ml
• Range
  5 µg/ml - 1000 µg/ml
• 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 ELISA more details

Properties

• 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
  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Mitochondrial markers

  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Cytochromes

  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Other Metabolism Assay

  • Signal Transduction
  • Metabolism
  • Mitochondrial

Applications

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

• 

  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)
  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)
  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

Protocol Booklet