Product nameAconitase Assay Kit
See all Aconitase kits
Sample typeCell Lysate, Tissue Lysate
Assay typeEnzyme activity
Assay time0h 40m
Aconitase Assay Kit ab83459 is a highly sensitive, simple, direct and HTS-ready colorimetric assay for measuring Aconitase activity in biological samples.
In the aconitase assay protocol, 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 max absorbance at 450nm.
Aconitase assay protocol summary:
- activate aconitase in samples by adding cysteine HCl and (NH4)2Fe(SO4)2
- add samples and standards to wells
- add reaction mix and incubate for 30-60 min
- analyze with a microplate reader
Aconitase (aconitate hydratase; EC 22.214.171.124) 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.
Storage instructionsStore at +4°C. Please refer to protocols.
Components Identifier 100 tests (NH4)2Fe(SO4)2 (Lyophilised) Brown/NM 1 vial Assay Buffer WM 1 x 25ml Cysteine (Lyophilised) Red 1 vial Developer (Lyophilised) Purple 1 vial Enzyme Mix Green 1 x 600µl Isocitrate Standard (100 mM) Yellow 1 x 100µl Substrate (lyophilized) Blue 1 vial
RelevanceAconitase (aconitate hydratase; EC 126.96.36.199) 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 prooxidants has been interpreted as a measure of oxidative damage.
Cellular localizationACO1: Cytoplasmic ACO2: Mitochondrial
- ACO 1
- ACO 2
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.
Mitochondrial autophagic profiles in ON vulnerable to secondary degeneration +/- R/NIR-IT. Accumulations of mitochondrial autophagic profiles (red arrows) were seen especially at day 1 after PT (A, B). Occasionally, larger features reminiscent of autophagosomes were observed (A, black arrow) and parts of the nerve appeared highly abnormal (*, B).
30 minute Aconitase sample test using ab83459.
Isocitrate standard curve using ab83459.
This product has been referenced in:
- Romsang A et al. Pseudomonas aeruginosa nfuA: Gene regulation and its physiological roles in sustaining growth under stress and anaerobic conditions and maintaining bacterial virulence. PLoS One 13:e0202151 (2018). Read more (PubMed: 30092083) »
- Langley M et al. Mito-Apocynin Prevents Mitochondrial Dysfunction, Microglial Activation, Oxidative Damage, and Progressive Neurodegeneration in MitoPark Transgenic Mice. Antioxid Redox Signal 27:1048-1066 (2017). Read more (PubMed: 28375739) »