Abcam's Aconitase Assay 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 λmax at 450nm.
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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.
Components | Identifier | 100 tests |
---|---|---|
(NH4)2Fe(SO4)2 (Lyophilised) | Brown/NM | 1 vial |
Assay Buffer | WM | 1 x 30ml |
Cysteine (Lyophilised) | Red | 1 vial |
Developer (Lyophilised) | Purple | 1 vial |
Enzyme Mix | Green | 1 x 200µl |
Isocitrate Standard (100 mM) | Yellow | 1 vial |
Substrate (lyophilized) | Blue | 1 vial |
Our Abpromise guarantee covers the use of ab83459 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
---|---|---|
Functional Studies | Use at an assay dependent dilution. |
Aconitase measured in mouse tissue lysates showing quantity (mU) per mg of tested sample.
Protein concentration for samples varied from 1.5 mg/mL to 10 mg/mL. Samples were diluted 1-27 fold.
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