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Complex IV Human Enzyme Activity Microplate Assay Kit ab109909 is used to determine the activity of cytochrome c oxidase in a human sample with greater speed and simplicity.


Images

Functional Studies - Complex IV Human Enzyme Activity Microplate Assay Kit (AB109909), expandable thumbnail
  • Functional Studies - Complex IV Human Enzyme Activity Microplate Assay Kit (AB109909), expandable thumbnail

Publications

Key facts

Detection method

Colorimetric

Sample types

Cell culture extracts, Tissue

Assay type

Enzyme activity

Reactive species

Cow, Human, Pig

Target data

Function

Component of the cytochrome c oxidase, the last enzyme in the mitochondrial electron transport chain which drives oxidative phosphorylation. The respiratory chain contains 3 multisubunit complexes succinate dehydrogenase (complex II, CII), ubiquinol-cytochrome c oxidoreductase (cytochrome b-c1 complex, complex III, CIII) and cytochrome c oxidase (complex IV, CIV), that cooperate to transfer electrons derived from NADH and succinate to molecular oxygen, creating an electrochemical gradient over the inner membrane that drives transmembrane transport and the ATP synthase. Cytochrome c oxidase is the component of the respiratory chain that catalyzes the reduction of oxygen to water. Electrons originating from reduced cytochrome c in the intermembrane space (IMS) are transferred via the dinuclear copper A center (CU(A)) of subunit 2 and heme A of subunit 1 to the active site in subunit 1, a binuclear center (BNC) formed by heme A3 and copper B (CU(B)). The BNC reduces molecular oxygen to 2 water molecules using 4 electrons from cytochrome c in the IMS and 4 protons from the mitochondrial matrix.

Alternative names

Recommended products

Complex IV Human Enzyme Activity Microplate Assay Kit ab109909 is used to determine the activity of cytochrome c oxidase in a human sample with greater speed and simplicity.

Key facts

Detection method

Colorimetric

Sample types

Cell culture extracts, Tissue

Assay type

Enzyme activity

Reactive species

Cow, Human, Pig

Assay Platform

Microplate reader

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

Multi

Appropriate long-term storage conditions

Multi

Storage information

Please refer to protocols

Notes

The Complex IV Human Enzyme Activity Microplate Assay Kit is used to determine the activity of cytochrome c oxidase in a human sample with greater 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.

Store Tube 1, detergent, and microplate at 4°C – DO NOT FREEZE. Store Reagent C at -20°C or at -80°C for long-term storage.

**Range of complex IV / cytochrome c oxidase assay kits** Biochemical assay - Immunocapture with biochemical assay (plate-based)*** - and (this kit) *** Most popular assay format Immunocapture with biochemical assay (dipstick) - and Immunocapture with biochemical assay and ELISA - 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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Associated diseases and disorders

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.

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2 product images

  • Functional Studies - Complex IV Human Enzyme Activity Microplate Assay Kit (ab109909), expandable thumbnail

    Functional Studies - Complex IV Human Enzyme Activity Microplate Assay Kit (ab109909)

    Figure 1. To determine the activity in the sample, calculate the slope by using microplate software or by manual calculations using one of the two methods shown. Compare the sample rate with the rate of the control (normal) sample and with the rate of the null (background) to get the relative Complex IV activity. (A)The rate is determined by calculating the gradient of the initial slope over the linear region. (B) The rate is determined by calculating the slope between two points within the linear region.

  • Functional Studies - Complex IV Human Enzyme Activity Microplate Assay Kit (ab109909), expandable thumbnail

    Functional Studies - Complex IV Human Enzyme Activity Microplate Assay Kit (ab109909)

    Abcam's enzyme activity assays apply a novel approach, whereby target enzymes are first immunocaptured from tissue or cell samples before subsequent functional analysis. All of our ELISA kits utilize highly validated monoclonal antibodies and proprietary buffers, which are able to capture even very large enzyme complexes in their fully-intact, functionally-active states.

    Capture antibodies are pre-coated in the wells of premium Nunc MaxiSorp™ modular microplates, which can be broken into 8-well strips. After the target has been immobilized in the well, substrate is added, and enzyme activity is analyzed by measuring the change in absorbance of either the substrate or the product of the reaction (depending upon which enzyme is being analyzed). By analyzing the enzyme's activity in an isolated context, outside of the cell and free from any other variables, an accurate measurement of the enzyme's functional state can be understood.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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