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
This product is manufactured by BioVision, an Abcam company and was previously called K716 Aconitase Activity Colorimetric Assay Kit. K716-100 is the same size as the 100 test size of ab83459.
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.
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Platform
Microplate reader
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 100 tests Ferrous Ammonium Sulfate 1 vial Assay Buffer VII 1 x 25ml Cysteine Standard 1 vial Developer Solution II 1 vial Enzyme Mix VII 1 x 600µl Isocitrate Standard 1 x 100µl Substrate XI 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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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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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).
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30 minute Aconitase sample test using ab83459.
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Isocitrate standard curve using ab83459.
Datasheets and documents
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SDS download
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Datasheet download
References (15)
ab83459 has been referenced in 15 publications.
- Álvarez-Córdoba M et al. Down regulation of the expression of mitochondrial phosphopantetheinyl-proteins in pantothenate kinase-associated neurodegeneration: pathophysiological consequences and therapeutic perspectives. Orphanet J Rare Dis 16:201 (2021). PubMed: 33952316
- Yao F et al. Iron regulatory protein 1 promotes ferroptosis by sustaining cellular iron homeostasis in melanoma. Oncol Lett 22:657 (2021). PubMed: 34386079
- Ghaisas S et al. Chronic Manganese Exposure and the Enteric Nervous System: An in Vitro and Mouse in Vivo Study. Environ Health Perspect 129:87005 (2021). PubMed: 34410835
- Flockhart M et al. Excessive exercise training causes mitochondrial functional impairment and decreases glucose tolerance in healthy volunteers. Cell Metab 33:957-970.e6 (2021). PubMed: 33740420
- Lombardi M et al. Mitochondrial Energetics and Ca2+-Activated ATPase in Obstructive Hypertrophic Cardiomyopathy. J Clin Med 9:N/A (2020). PubMed: 32527005