MitoSciences (MS241)

Complex II Enzyme Activity Microplate Assay Kit (ab109908)

Overview

  • Product nameComplex II Enzyme Activity Microplate Assay Kit
    See all Complex II kits
  • Detection methodColorimetric
  • Assay typeEnzyme activity
  • Assay durationMultiple steps standard assay
  • 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 and DCPIP should be stored at -80°C. Lipid Mix should be stored at -20°C. All other components should be stored at 4°C.

  • Tested applicationsFunctional Studiesmore details
  • PlatformMicroplate

Properties

Applications

Our Abpromise guarantee covers the use of ab109908 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
Functional Studies Use at an assay dependent dilution.

Complex II Enzyme Activity Microplate Assay Kit 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 for Complex II Enzyme Activity Microplate Assay Kit (ab109908)

This product has been referenced in:
  • Thongchote K  et al. Running exercise alleviates trabecular bone loss and osteopenia in hemizygous ß-globin knockout thalassemic mice. Am J Physiol Endocrinol Metab 306:E1406-17 (2014). Read more (PubMed: 24780610) »
  • Alfaradhi MZ  et al. Oxidative stress and altered lipid homeostasis in the programming of offspring fatty liver by maternal obesity. Am J Physiol Regul Integr Comp Physiol 307:R26-34 (2014). Functional Studies ; Mouse . Read more (PubMed: 24789994) »

See all 13 Publications for this product

Product Wall

Thank you very much for your call today and for your patience while I have been in touch with the lab scientists regarding this enquiry.

If you're using a tissue homogenate or whole mitochondria sample, before extraction with the kit’s ...

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Thank you for your inquiry.


The assays can be optimized by cell number, but since all cells are different sizes we use total protein mass.


In general large cells such as fibroblast, HeLa etc are about 0.25 mg/million. So...

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According to the literature, malonate inhibits complex II with an IC50 of ˜40 μM (PMCID: PMC2507763). Please let me know if you have any further questions!

Thank you for contacting us.

Yes, you should keep samples on ice, and also add a protease inhibitor to the detergent.

Yes TTFA can be added and should decrease the activity of Complex II.
However TTFA is not as specific/effect...

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Pipette up and down cells several time for complete lysis and then determine the protein concentration using BCA or Bradford method. Once protein concentration is known then please follow rest of the procedure.

20 million of average sized cel...

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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.
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Mr. Amit Agarwal

Verified customer

Submitted Jul 24 2013

My guess is that the levels of OXPHOS proteins in whole blood are below the level of detection for activity assays. The activity assays for the most part require higher loading than the sandwich ELISA assays. We have tested Complex IV Quantity with who...

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Es werden keinen zusätzlichen Proteinaseinhibitoren für das Kit (ab109908, Complex II Enzyme Activity Microplate Assay Kit) benötigt.

Wir empfehlen während der Aufbereitung der Proben (bevor das Kit benutzt wird;...

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We use DCPIP as a the detector in this kit. DCPIP is an enzyme-catalyzed oxidation electron acceptor that is blue in its oxidized form and colorless in its reduced form. A peak in the absorbance is observed at 600 nm, however you will still be able to ...

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