Description
Key facts
Detection method:Colorimetric,
Sample types:Inhibitor compounds,
Assay type:Quantitative,
Assay time:5h,
Assay Platform:Microplate reader
Product details:
MitoTox Complex I OXPHOS Activity Assay (ab109903) is designed for testing the direct inhibitory effect of compounds on Complex I activity in only 5 hours. Complex I extracted from the provided bovine heart mitochondria (a rich source of Complex I) is immunocaptured by specific antibodies on the plate. Complex I activity can be observed as decrease in absorbance at OD 340 nm. The intra-assay and inter-assay variation of this assay are both < 10%.
Inhibitory effects of compounds on Complex I 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 (IC
) format, where two compounds known to affect Complex I 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.
Store Phospholipids, Bovine heart mitochondria, Ubiquinone and Activity Buffer at -80°C. Store all other components at 4°C.
Properties and Storage Information:
Shipped at conditions-Dry Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Complex I also known as NADH dehydrogenase or NADH:ubiquinone oxidoreductase is a large enzyme complex with a molecular mass of approximately 1000 kDa. It is expressed in the inner mitochondrial membrane of eukaryotic cells. As the first enzyme in the mitochondrial respiratory chain Complex I plays a critical mechanical role in cellular respiration. It transfers electrons from NADH to ubiquinone coupled with the translocation of protons across the inner mitochondrial membrane contributing to the generation of a proton gradient used to produce ATP.
Biological function summary
Complex I acts as an integral component of the mitochondrial respiratory chain which is a series of protein complexes involved in cellular energy production. As part of this complex system Complex I is essential for effective oxidative phosphorylation. Its activity is assessed using protein activity assays including immunocapture or complex activity assays and microplate assays. Complex I activity influences the overall efficiency of ATP production affecting energy-dependent cellular processes.
Pathways
Complex I functions within the electron transport chain one of the major pathways in cellular respiration. This pathway is vital for ATP synthesis providing the energy currency required by cells. Complex I works closely with other electron transport chain complexes such as Complex II (succinate dehydrogenase complex) and Complex III (cytochrome c reductase) to drive oxidation-reduction reactions and maintain cellular metabolism.
Defects in Complex I are linked with mitochondrial diseases and neurodegenerative disorders such as Leigh syndrome and Parkinson's disease. Mutations in Complex I subunits disrupt normal electron transport and ATP production leading to increased oxidative stress and neuronal cell damage. Other mitochondrial proteins such as cytochrome c oxidase are also implicated in these conditions emphasizing the interconnected nature of mitochondrial dysfunction in disease progression.
