Complex II Enzyme Activity Microplate Assay Kit (ab109908)

Overview

• Product nameComplex II Enzyme Activity Microplate Assay KitSee all Complex II kits ...
• Assay typeEnzyme activity
• 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 Studies more details

Properties

• 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
  • Metabolism
  • Pathways and Processes
  • Mitochondrial Metabolism
  • Mitochondrial markers

  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Other Metabolism Assay

  • Signal Transduction
  • Metabolism
  • Mitochondrial

Applications

Complex II Enzyme Activity Microplate Assay Kit images

• 

  Functional Studies - Complex II Enzyme Activity Microplate Assay Kit (ab109908)
  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.
• 

  - Complex II Enzyme Activity Microplate Assay Kit (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.
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  - Complex II Enzyme Activity Microplate Assay Kit (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.

Protocols

Protocol Booklet