Description
Key facts
Detection method:Colorimetric,
Sample types:Inhibitor compounds,
Reacts with:Rat, Cow, Human,
Assay type:Direct,
Assay time:4h,
Assay Platform:Microplate reader
Product details:
MitoTox™ Complex V OXPHOS Activity Assay Kit (ab109907) is designed for testing the direct inhibitory effect of compounds on Complex V activity in only 4 hours. Complex V extracted from the provided bovine heart mitochondria (a rich source of Complex V) is immunocaptured by specific antibodies on the plate. Complex V 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 V 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 and Activity Buffer at -80°C. Store all other components at 4°C.**Related products**Review the , or the full 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.
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 V commonly called ATP synthase plays an important role in cellular energy production. This enzyme complex exists in the inner mitochondrial membrane and is approximately 600 kDa in size. Complex V facilitates the conversion of ADP and inorganic phosphate into ATP during oxidative phosphorylation. It consists of multiple subunits including Fo and F1 components where Fo functions as a proton channel while F1 performs ATP synthesis. Complex V is ubiquitously expressed across various tissues reflecting its essential role in energy generation.
Biological function summary
Complex V is important for maintaining cellular energy homeostasis by generating ATP the cell's energy currency. It operates as part of the mitochondrial respiratory chain complex essential for efficient energy transfer and storage. This complex plays a significant role in cellular metabolism by linking the electrochemical gradient formed by protons across the inner mitochondrial membrane to the mechanical rotation of its subunits leading to ATP formation. The ATP synthase products are imperative for processes demanding high energy such as muscle contraction and biosynthetic pathways.
Pathways
Complex V integrates into major metabolic routes primarily oxidative phosphorylation and the broader mitochondrial electron transport chain. It works in conjunction with complexes I to IV facilitating energy conversion via a proton gradient and ATP generation. Complex V's role complements proteins such as cytochrome c and ubiquinone which shuttle electrons within the electron transport chain ensuring a seamless energy production process essential for aerobic respiration.
Complex V dysfunction can cause mitochondrial diseases and disorders related to energy metabolism including Leigh syndrome and mitochondrial myopathy. These conditions often result from mutations in the ATP synthase subunits leading to impaired ATP production and altered cellular functions. Additional related proteins in these diseases include NADH dehydrogenase and succinate dehydrogenase which when dysfunctional further contribute to defects in the electron transport chain compounding the energy production challenges in affected individuals.
