Description
Key facts
Detection method:Colorimetric,
Sample types:Cell culture extracts, Tissue,
Reacts with:Mouse, Rat,
Assay type:Enzyme activity (quantitative),
Assay time:6h,
Assay Platform:Microplate reader
Product details:
Complex IV Rodent Enzyme Activity Microplate Assay Kit (ab109911) is used to determine the activity of cytochrome c oxidase in a mouse sample with speed and simplicity.
The COX enzyme is immunocaptured within the wells of the microplate and activity is determined colorimetrically by following the oxidation of reduced cytochrome c by the absorbance change at 550 nm.
Included in this kit for performance of the activity assay are buffer, detergent, substrate, and 96-well microplate with monoclonal antibody pre-bound to the wells of the plate, allowing for a stream-lined assay.
Complex IV assay protocol summary
- add samples to wells to capture enzyme and incubate for 3 hrs
- wash wells
- add reaction mix
- analyze with microplate reader in kinetic mode for 120 min
Buffer, detergent, and microplate should be stored at 4°C, Reagent C (reduced cytochrome c) should be stored at -80°C.
Range of complex IV / cytochrome c oxidase assay kits
Immunocapture with biochemical assay (plate-based) ab109911 (this kit) and
ab109909
Immunocapture with biochemical assay (dipstick)
ab109878
ab109876
Immunocapture with biochemical assay and ELISA
ab109910
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.
