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.

  • Notes

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

    Related products

    Review the mitochondrial assay guide, or the full metabolism assay guide to learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.

  • Platform
    Microplate reader

Properties

Images

  • 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.

    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.
  • 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 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.
  • 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.

Protocols

References

This product has been referenced in:
See all 39 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

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