Key features and details
- Assay type: Direct
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 4 hr
Product nameMitoTox™ Complex II OXPHOS Activity Assay Kit
See all Complex II kits
Assay time4h 00m
Species reactivityReacts with: Mouse, Cow, Human
MitoTox™ Complex II OXPHOS Activity Assay Kit (ab109904) is designed for testing the direct inhibitory effect of compounds on Complex II activity in only 4 hours. Complex II extracted from the provided bovine heart mitochondria (a rich source of Complex II) is immunocaptured by specific antibodies on the plate. Complex II activity can be observed as decrease in absorbance at OD 600 nm. The intra-assay and inter-assay variation of this assay are both <15%.
Inhibitory effects of compounds on Complex II activity can be tested in two different ways: 1. Screening format, where up to 23 compounds can be tested at a single concentration in triplicate; 2. Dose response (IC50) format, where two compounds known to affect Complex II activity can be tested at 11 different data points in triplicate.
Testing for mitochondrial function has become a key aspect of drug discovery. Mitochondria can be affected by drug treatment, resulting into cardio- and hepatotoxic side effects that can lead to drug withdrawal from the market. Therefore, there is increasing emphasis on testing the impact on mitochondria early on in the drug development process to reduce failure rates during preclinical and clinical phases.
Please store Succinate, Ubiquinone 2, Bovine Heart Mitochondria, DCPIP at -80°C and all other components at 4°C.
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.
Storage instructionsPlease refer to protocols.
Components 96 tests Detergent 1 x 1ml 12-channel reagent reservoirs 2 units 20X Buffer 1 x 5ml Bovine heart mitochondria 2 x 360µl Complex II Activity Buffer 1 x 25ml DCPIP/DCIP 1 x 250µl Pre-coated 96-well microplate 1 x 96 tests Succinate 1 x 500µl Ubiquinone 2 1 x 60µl
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
Typical dose response curve for TTFA (2-thenoyltrifluoracetone). Assay was performed following the Dose Response Assay Procedure using TTFA, a well known Complex II inhibitor. TTFA was prepared in DMSO to generate a 100 mM stock. Starting with a 500 µM final concentration in well, 1:2 serial dilutions of TTFA were generated.
ab109904 has been referenced in 3 publications.
- Subedi A et al. A novel inhibitor of tumorspheres reveals the activation of the serine biosynthetic pathway upon mitochondrial inhibition. FEBS Lett 593:763-776 (2019). PubMed: 30874300
- Landry GM et al. Diglycolic acid inhibits succinate dehydrogenase activity in human proximal tubule cells leading to mitochondrial dysfunction and cell death. Toxicol Lett 221:176-84 (2013). PubMed: 23827505
- Lai K et al. Integrated Compound Profiling Screens Identify the Mitochondrial Electron Transport Chain as the Molecular Target of the Natural Products Manassantin, Sesquicillin, and Arctigenin. ACS Chem Biol N/A:N/A (2012). PubMed: 23138533