Aconitase Assay Kit (ab83459)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 40 min
- Sample type: Cell Lysate, Tissue Lysate
Overview
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Product name
Aconitase Assay Kit
See all Aconitase kits -
Detection method
Colorimetric -
Sample type
Cell Lysate, Tissue Lysate -
Assay type
Enzyme activity -
Assay time
0h 40m -
Product overview
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 -
Notes
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.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate reader
Properties
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Storage instructions
Store 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 -
Research areas
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Relevance
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 prooxidants has been interpreted as a measure of oxidative damage. -
Cellular localization
ACO1: Cytoplasmic ACO2: Mitochondrial -
Alternative names
- ACO 1
- ACO 2
- ACO1
see all
Images
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Functional Studies - Aconitase Assay Kit (ab83459)
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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Functional Studies - Aconitase Assay Kit (ab83459) Cummins, Nadia et al., BMC neuroscience?vol. 14 98., Fig 6, doi:10.1186/1471-2202-14-98Mitochondrial 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).
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Functional Studies - Aconitase Assay Kit (ab83459)30 minute Aconitase sample test using ab83459. -
Functional Studies - Aconitase Assay Kit (ab83459)Isocitrate standard curve using ab83459.
