Human Succinate Dehydrogenase ELISA Kit (Complex II) Profiling ELISA Kit (ab124536)

Overview

  • Product nameHuman Succinate Dehydrogenase ELISA Kit (Complex II) Profiling ELISA Kit
    See all Complex II kits
  • Detection methodColorimetric
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Overall 5 3.9%
    Inter-assay
    Sample n Mean SD CV%
    Overall 3 15%
  • Tests
    1 x 96 test
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay typeSandwich (qualitative)
  • Sensitivity
    50 µg/ml
  • Range
    12 µg/ml - 1000 µg/ml
  • Assay durationMultiple steps standard assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Mouse, Rat
  • Product overview

    Abcam’s Succinate dehydrogenase (Complex II) Profiling in vitro ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for comparison of Succinate dehydrogenase levels in Human cell and tissue lysates.

    The assay employs a Human succinate dehydrogenase complex specific capture coated onto microplate well plate strips. Samples are pipetted into the wells and succinate dehydrogenase present in the sample is bound to the wells by the immobilized antibody. The wells are washed and an anti-succinate dehydrogenase 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 succinate dehydrogenase (Complex II) 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

    Succinate dehydrogenase (Complex II) is the second enzyme of the oxidative phosphorylation (OXPHOS) system within the mitochondrial inner membrane. Succinate dehydrogenase is a large protein complex of 140,000 MW made up of 4 different subunits (SDH, SDB, SDHC, and SDHD). The enzyme forms a dimer in the mitochondrial inner membrane. Succinate dehydrogenase is the only enzyme of the OXPHOS system containing no mitochondrial DNA (mtDNA) encoded subunit components. Therefore all four subunits are encoded by nuclear genomic DNA, made in the cytosol, and translocated into the organelle for assembly at the inner membrane. The enzyme complex catalyses electron entry from succinate via a flavin adenine dinucleotide (FAD), three iron sulfur clusters and b-type cytochrome. Succinate dehydrogenase also participates in the citric acid cycle producing fumarate for the next step in the cycle - hydration by fumarase.

  • Tested applicationsSuitable for: Sandwich ELISAmore details
  • PlatformMicroplate

Properties

Applications

Our Abpromise guarantee covers the use of ab124536 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.

Human Succinate Dehydrogenase ELISA Kit (Complex II) Profiling ELISA Kit images

  • Example control sample curve of serially titrated HepG2 lysate in the working range of the assay.
  • Example control sample curve of serially titrated human heart tissue homogenate in the working range of the assay.
  • Example control sample curve of serially titrated HL60 lysate 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, 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.
  • 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.

Protocols

References for Human Succinate Dehydrogenase ELISA Kit (Complex II) Profiling ELISA Kit (ab124536)

ab124536 has not yet been referenced specifically in any publications.

Product Wall



El buffer utilizado consiste en 1.5% de lauryl maltoside. Este detergente es compatible con la mayoría de kits ELISA para medir actividad del proceso de fosforilación oxidativa, excluyendo la medida del kit para determinar la ac...

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