Complex II Enzyme Activity Microplate Assay Kit (ab109908)
- Product nameComplex II Enzyme Activity Microplate Assay KitSee all Complex II kits ...
- Detection methodColorimetric
- Assay typeEnzyme activity
- Assay durationMultiple steps standard assay
- Species reactivityReacts 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.
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 Studies more details
- Storage instructionsPlease refer to protocols.
Components 96 tests 10X Blocking Solution 1 x 5ml 10X Detergent 2 x 1ml 20X Buffer 1 x 15ml Complex II Activity Buffer 1 x 25ml DCPIP 1 x 250µl Lipid Mix 1 x 6ml Pre-coated 96-well microplate (12 strips) 1 unit Succinate 1 x 500µl Ubiquinone 2 1 x 60µl
- Research Areas
- RelevanceComplex II is also called succinate ubiquinone oxidoreductase or more commonly succinate dehydrogenase complex. This complex is composed of four nuclear encoded subunits and contains a flavin (FAD), non-heme iron centers and a b-type cytochrome as prosthetic groups. It is both a component of the electron transport chain and an enzyme of the Krebs cycle. Complex II deficiencies are seen in OXPHOS genetic disease and found in a type of cancer called paraganglioma.
- Succinate coenzyme Q reductase
- Succinate dehydrogenase
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.
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.
References for Complex II Enzyme Activity Microplate Assay Kit (ab109908)
This product has been referenced in:
- Kumarasamy S et al. Construction of two novel reciprocal conplastic rat strains and characterization of cardiac mitochondria. Am J Physiol Heart Circ Physiol 304:H22-32 (2013). Rat . Read more (PubMed: 23125210) »
- Li C et al. PI3K/AKT signaling regulates bioenergetics in immortalized hepatocytes. Free Radic Biol Med 60C:29-40 (2013). Functional Studies . Read more (PubMed: 23376468) »