Aconitase (aconitate hydratase; EC 4.2.1.3) is an iron-sulfur protein containing an [Fe4S4]2+ cluster that catalyzes the stereospecific isomerization of citrate to isocitrate via cis-aconitate in the tricarboxylic acid cycle, a non-redox-active process. Tissue contains two aconitases, a mitochondrial (m-) and a cytosolic (c-) aconitase. They are related, but distinctly different enzymes and are coded for on different chromosomes. Loss of aconitase activity in cells or other biological samples treated with pro-oxidants has been interpreted as a measure of oxidative damage. Aconitase BioAssay Kit is a highly sensitive, simple, direct and HTS-ready colorimetric assay for measuring Aconitase activity in biological samples. In the assay, citrate is converted by aconitase into isocitrate, which is further processed resulting in a product that converts a nearly colorless probe into an intensely colored form with a Lmax at 450nm. the readings are within the linear range.Kit Components:1. Assay Buffer: 1x 30ml2. Substrate: lyophilized3. Developer: lyophilized 4. Enzyme Mix: 1x200ul) 5. Cysteine: lyophilized 6. (NH 4)2Fe(SO4)2: lyophilized 7. Isocitrate Standard (10umol): lyophilized Storage and Stability:Store kit at -20°C. Protect from light. Allow Assay Buffer to warm to room temperature before use. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
