Description
Key facts
Detection method:Colorimetric,
Sample types:Tissue Lysate, Cell Lysate,
Assay type:Enzyme activity,
Assay time:40m,
Assay Platform:Microplate reader
Product details:
Aconitase Assay Kit ab83459 is a highly sensitive, simple, direct and HTS-ready colorimetric assay for measuring Aconitase activity in biological samples.
Aconitase assay principle
In the aconitase assay protocol, citrate is converted by aconitase into isocitrate. Isocitrate dehydrogenase, then acts on isocitrate and NADP, in a coupled reaction that converts a probe into an colored form with a max absorbance at 450nm.
Aconitase assay protocol summary
- activate aconitase in samples by adding cysteine HCl and (NH
- add samples and standards to wells
- add reaction mix and incubate for 30-60 min
- analyze with a microplate reader
Related Aconitase assay kits
< br/>For an alternative aconitase assay method, based on measurement of absorbance of the cis-aconitate intermediate formed during the action of aconitase, use Aconitase assay kit
ab109712
Other notes
This was previously called K716 Biovision Aconitase Activity Colorimetric Assay Kit. Biovision was acquired by Abcam in 2021
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.
REACH authorisation
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.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-+4°C, Appropriate long-term storage conditions-+4°C, Storage information-Please refer to protocols
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Aconitase also known as aconitate hydratase is an enzyme with a mass of approximately 82 kDa. It catalyzes the reversible isomerization of citrate to isocitrate via cis-aconitate in the tricarboxylic acid (TCA) cycle. Aconitase is expressed in the mitochondria where it functions as mitochondrial aconitase and in the cytoplasm in a slightly different form. The enzyme contains an iron-sulfur cluster that is important for its activity. Scientists often use aconitase kits to measure its activity in different samples.
Biological function summary
Mitochondrial aconitase plays an important role in energy production by facilitating the TCA cycle which is essential for ATP generation. The enzyme can also serve as an iron regulatory protein in the cytoplasm contributing to the regulation of iron metabolism. Aconitase exists as part of a larger complex with its iron-sulfur cluster playing a critical role in both its enzymatic and regulatory functions. By shifting between its enzymatic and regulatory roles aconitase helps in maintaining homeostasis within cells.
Pathways
Aconitase operates in the TCA cycle an important metabolic pathway for energy production. It participates in the metabolism of acetyl-CoA converting it into isocitrate through interactions with citrate. This key pathway integrates with glycolysis and aids in the oxidative phosphorylation process. Aconitase's function is related to proteins such as isocitrate dehydrogenase which further processes isocitrate in the TCA cycle.
Mutations and defects in aconitase can contribute to neurodegenerative diseases and iron-sulfur cluster deficiencies. A connection exists between aconitase activity and disorders like Friedreich's ataxia a condition featuring impaired iron-sulfur cluster biosynthesis. Ferritin a protein associated with iron storage links with aconitase through pathways involved in iron regulation suggesting potential therapeutic targets in disorders of iron metabolism.
