Description
Key facts
Detection method:Colorimetric,
Sample types:Cell culture extracts, Tissue,
Reacts with:Human,
Assay type:Enzyme activity,
Assay Platform:Microplate reader
Product details:
This rapid multiplexing microplate kit is used to determine both the activity and quantity of cytochrome c oxidase in a human sample with greater speed and simplicity. The ratio of the two is a measure of specific activity. The COX enzyme is immunocaptured within the wells of the microplate and the activity is determined by following the oxidation of reduced Cytochrome c colorimetrically by the absorbance change at 550 nm. Subsequently, in these same wells the quantity of the enzyme can then be determined by adding a COX specific detector antibody and the appropriate alkaline phophatase conjugated antibody; this phosphatase changes a substrate from colorless to yellow at 405 nm. This reaction takes place in a time dependent manner proportional to the amount of protein captured in the wells. Included in this kit for performance of the multiplexing assay are buffers, detergent, substrates, and 96-well microplate with monoclonal antibody pre-bound to the wells of the plate, allowing for a stream-lined assay.
**Range of complex IV / cytochrome c oxidase assay kits**Biochemical assay - Immunocapture with biochemical assay (plate-based)*** - and *** Most popular assay formatImmunocapture with biochemical assay (dipstick) - and Immunocapture with biochemical assay and ELISA - (this kit)ELISA - **Other 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.
