Description
Key facts
Detection method:Colorimetric,
Reacts with:Cow, Human,
Assay type:Direct,
Assay time:5h,
Assay Platform:Microplate reader
Product details:
MitoTox™ Complex IV OXPHOS Activity Assay Kit (ab109906) is designed for testing the direct inhibitory effect of compounds on Complex IV activity in only 5 hours. Complex IV extracted from the provided bovine heart mitochondria (a rich source of Complex IV) is immunocaptured by specific antibodies on the plate. Complex IV activity can be observed as decrease in absorbance at OD 550 nm. The intra-assay and inter-assay variation of this assay are both <10%.
Inhibitory effects of compounds on Complex IV 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 IV 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 Bovine Heart Mitochondria and Reagent C 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 IV also known as cytochrome c oxidase is an important component of the electron transport chain in mitochondria. This enzyme complex has significant mass approximately 204 kDa and operates primarily in the inner mitochondrial membrane. It catalyzes the transfer of electrons from cytochrome c to oxygen facilitating ATP generation. Complex IV is ubiquitously expressed in all tissues with higher expression in organs with high energy demands like heart and skeletal muscle. This enzyme is composed of multiple subunits encoded by both mitochondrial and nuclear DNA.
Biological function summary
Cytochrome c oxidase plays a critical role in cellular respiration. It forms part of the larger enzyme complex which also includes Complex I II and III. These complexes work together to create a proton gradient across the inner mitochondrial membrane essential for ATP synthesis via oxidative phosphorylation. This enzyme's activity is measured through assays like the cytochrome c oxidase assay which evaluates its function in various tissues. These assays help researchers understand how effectively electrons are being transferred and protons are driven across the membrane.
Pathways
Complex IV is integral to the oxidative phosphorylation pathway and the broader mitochondrial respiratory chain. It interacts closely with cytochrome c a small heme protein that shuttles electrons between Complex III (cytochrome c reductase) and Complex IV. Through these interactions the proton gradient is established enabling ATP synthase to convert chemical energy into usable cell energy. This process significantly impacts cellular metabolism and energy production influencing how efficiently cells function.
Complex IV dysfunction is associated with mitochondrial diseases and certain neurodegenerative disorders. For instance defects in cytochrome c oxidase lead to conditions like Leigh syndrome a severe neurological disorder. Also studies show that disruptions in the electron transport chain involving Complex IV relate closely to Alzheimer’s disease. Alterations in proteins like cytochrome c which work in tandem with Complex IV can exacerbate these conditions highlighting the importance of this target in understanding and potentially treating these diseases.
