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Human Cyt C Reductase ELISA Kit (Complex III) Profiling ELISA Kit (ab124537)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (1)References (5)

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Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)
  • Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)
  • Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)

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

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Overview

  • Product name

    Human Cyt C Reductase ELISA Kit (Complex III) Profiling ELISA Kit
    See all Complex III kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    Overall 8 4.7%
    Inter-assay
    Sample n Mean SD CV%
    Overall 6 19.7%
  • Sample type

    Tissue Extracts, Cell Lysate
  • Assay type

    Sandwich (qualitative)
  • Sensitivity

    4 µg/ml
  • Range

    6 µg/ml - 200 µg/ml
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    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.

  • 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.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store 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
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
  • Relevance

    Complex 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.
  • Alternative names

    • coenzyme Q : cytochrome c — oxidoreductase
    • cytochrome bc1 complex
  • Database links

    • Entrez Gene: 7384 Human
    • Entrez Gene: 7385 Human
    • Entrez Gene: 1537 Human
    • Entrez Gene: 7386 Human
    • Entrez Gene: 4519 Human
    • Entrez Gene: 7381 Human
    • Entrez Gene: 7388 Human
    • Entrez Gene: 27089 Human
    • Entrez Gene: 29796 Human
    • Entrez Gene: 10975 Human
    • SwissProt: P31930 Human
    • SwissProt: P22695 Human
    • SwissProt: P08574 Human
    • SwissProt: P47985 Human
    • SwissProt: P00156 Human
    • SwissProt: P07919 Human
    • SwissProt: P14927 Human
    • SwissProt: O14949 Human
    • SwissProt: Q9UDW1 Human
    • SwissProt: O14957 Human
    see all

Images

  • Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)
    Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)

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

  • Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)
    Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)
    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.
  • Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)
    Sandwich ELISA - Human Cyt C Reductase (Complex III) Profiling ELISA Kit (ab124537)
    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.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (5)

Publishing research using ab124537? Please let us know so that we can cite the reference in this datasheet.

ab124537 has been referenced in 5 publications.

  • 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

Customer reviews and Q&As

Show All Reviews Q&A
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Question

How can I analyze my data?





Read More

Abcam community

Verified customer

Asked on Nov 18 2014

Answer

For interpolation, using plate reader software is often best and easiest. Alternative to that would be Graphpad. Excel is poor at curve fitting and interpolation.

You can use color development rate or endpoint at any given time to construct the graph either manually or preferrably using the software.

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Nov 18 2014

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