人Cyt C Reductase ELISA试剂盒(Complex III) Profiling ELISA试剂盒(ab124537)

概述

  • 产品名称人Cyt C Reductase ELISA试剂盒(Complex III) Profiling ELISA试剂盒
    参阅全部 Complex III 试剂盒
  • 检测方法Colorimetric
  • 精确度
    批次内
    样品 n Mean SD CV%
    Overall 8 4.7%
    批次间
    样品 n Mean SD CV%
    Overall 6 19.7%
  • 测试
    1 x 96 test
  • 样品类型
    Tissue Extracts, Cell Lysate
  • 检测类型Sandwich (qualitative)
  • 灵敏度
    4 µg/ml
  • 范围
    6 µg/ml - 200 µg/ml
  • 实验步骤Multiple steps standard assay
  • 种属反应性
    与反应: Human
    不与反应: Mouse, Rat
  • 产品概述

    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.

  • 说明

    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.

  • 经测试应用适用于: Sandwich ELISAmore details
  • 平台Microplate

性能

应用

Our Abpromise guarantee covers the use of ab124537 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
Sandwich ELISA Use at an assay dependent concentration.

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

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

  • 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.
  • 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.

实验方案

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

This product has been referenced in:
  • Cherry AB  et al. Induced Pluripotent Stem Cells with a Pathological Mitochondrial DNA Deletion. Stem Cells N/A:N/A (2013). ELISA ; Human . Read more (PubMed: 23400930) »

See 1 Publication for this product

Product Wall

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