Cytochrome c Oxidase (EC 1.9.3.1) or Complex IV is the fourth complex of the Electron Transport Chain located in the mitochondrial (or bacterial) membrane. It provides energy to the cell by coupling electron transport through the Cytochrome c chain with the process of oxidative phosphorylation. Complex IV contains 13 different subunits encoded by both mitochondrial DNA and nuclear DNA. It receives an electron from each of the four Cytochrome c molecules, and transfers it to one oxygen molecule, converting it into two molecules of water. In this process, it also binds to four proton molecules and translocates them across the membrane to establish electrochemical gradient, which is utilized for the synthesis of ATP.
Sample Type:
Purified mitochondria and cells/tissue extracts.
Intended Use:
The kit is useful for mitochondrial respiration studies, assembly of the complexes, mitochondria outer membrane integrity.
Test Principle:
Cytochrome Oxidase Activity BioAssay(TM) Kit is simple, fast and high-throughput adaptable. This assay kit can be used for purified mitochondria or tissue extracts containing mitochondria. The activity of the enzyme is determined colorimetrically by following the oxidation of reduced Cytochrome c as an absorbance decrease at 550nm.
Kit Components:
Cytochrome Oxidase Assay Buffer, 1x25ml
Enzyme Dilution Buffer, 1x8ml
1M DTT, 1x1ml
Cytochrome c, 2x vials
96-well Plate, 1x plate
Storage and Stability:
Store powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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