Overview

  • Product name
    Complex II Enzyme Activity Microplate Assay Kit
    See all Complex II kits
  • Detection method
    Colorimetric
  • Sample type
    Cell Lysate, Tissue Lysate, Purified mitochondria
  • Assay type
    Enzyme activity
  • Assay time
    3h 00m
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
  • Product overview

    Complex II Enzyme Activity Microplate Assay Kit is designed for determining the Complex II activity in a human, mouse, rat or bovine sample. Each of the 96 wells in the kit has been coated with an anti-Complex II monoclonal antibody (mAb) which purifies the enzyme from a complex sample such as mitochondria, tissue homogenate or cell lysate. After this in-well purification the production of ubiquinol by the enzyme is coupled to the reduction of the dye DCPIP (2,6-diclorophenolindophenol) and a decreases in its absorbance at 600 nm, which in turn recycles the substrate ubiquinone.


    Review our Metabolism Assay Guide to learn about assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also about how to assay metabolic function in live cells using your plate reader.

  • Notes

    Succinate, Ubiquinone 2, DCPIP and Lipid?Phospholipd Mix should be stored at -80°C. All other components should be stored at 4°C.

  • Platform
    Microplate reader

Properties

Images

  • Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. All of our ELISA kits utilize highly validated monoclonal antibodies and proprietary buffers, which are able to capture even very large enzyme complexes in their fully-intact, functionally-active states.

    Capture antibodies are pre-coated in the wells of premium Nunc MaxiSorp™ modular microplates, which can be broken into 8-well strips. After the target has been immobilized in the well, substrate is added, and enzyme activity is analyzed by measuring the change in absorbance of either the substrate or the product of the reaction (depending upon which enzyme is being analyzed). By analyzing the enzyme's activity in an isolated context, outside of the cell and free from any other variables, an accurate measurement of the enzyme's functional state can be understood.
  • Figure 1. Example of raw data. Note the lag period before activity. Also note the activity of mitochondria (BHM, bovine heart mitochondria) is higher than whole cell lysate (HepG2, human hepatoblastoma) and the reaction ends at >1600 seconds because the substrates are used up.
  • Figure 2. This assay is compatible with different sample types such as mitochondria, tissue or cell lysates and in multiple species including human and rodent samples. Typical linear range data are shown for ab109908.

Protocols

References

This product has been referenced in:
  • Tetri LH  et al. Sex-Specific Skeletal Muscle Fatigability and Decreased Mitochondrial Oxidative Capacity in Adult Rats Exposed to Postnatal Hyperoxia. Front Physiol 9:326 (2018). Read more (PubMed: 29651255) »
  • Andreux PA  et al. Mitochondrial function is impaired in the skeletal muscle of pre-frail elderly. Sci Rep 8:8548 (2018). Read more (PubMed: 29867098) »
See all 35 Publications for this product

Customer reviews and Q&As

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1-2 of 2 Abreviews

Abreviews
Easy to follow instruction and standardize using cell lysate. Good sensitivity with microplate reader

Abcam user community

Verified customer

Submitted Feb 27 2018

30ug Lung mitochondrion samples from mice in buffer containing 250 mM sucrose, 20 mM HEPES, 1 mM EDTA, 1 mM EGTA, 0.25% protease inhibitor, and 0.5% BSA, pH 7.4 were used for the assay.

Dr. Amit Agarwal

Verified customer

Submitted Jul 24 2013

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

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