Description
Key facts
Detection method:Colorimetric,
Sample types:Urine, Plasma, Other biological fluids,
Assay type:Quantitative,
Sensitivity:< 25 µM,
Assay time:25m,
Assay Platform:Microplate reader
Product details:
Acetoacetate Assay Kit (Colorimetric) ab180875 adapts the traditional qualitative assay for acetoacetate using dipsticks, and sodium nitroferricyanide as a chromophore, to provide an assay optimized for use with plate readers that can be used to quantitate endogenous levels of AcAc in human urine, plasma and other biological fluids.
Acetoacetate assay principle
In the nitroferricyanide method acetoacetate assay, AcAc reacts with sodium nitroferricyanide to generate a colored product that can be measured at 550 nm. The reaction is specific for AcAc and does not detect 3-β-hydroxybutyrate. The assay kit can detect samples containing acetoacetate as low as 25 μM.
Acetoacetate assay protocol summary
- add samples and standards to wells
- add reaction mix
- analyze with a microplate reader in kinetic mode for 10-15 min
Related and recommended products
Acetoacetate (AcAc), a β-ketoacid, is one of the three ketone bodies and is formed via condensation of two molecules of acetyl-CoA in liver mitochondria. AcAc can be enzymatically reduced to 3-β-hydroxybutyrate (β-HB), or decarboxylated producing acetone ((CH
CO). Ketone bodies [βHB: 78%; AcAc: 20% & (CH
CO: 2%] are mainly used as an alternative energy source when glucose cannot be delivered to the system. Excessive concentration of ketone bodies (ketoacidosis) is observed in patients with Type I diabetes, severe starvation or alcoholism. Traditionally, AcAc levels have been qualitatively detected using dipsticks that use sodium nitroferricyanide as a chromophore
This assay kit is used to measure the acetoacetate element of ketone bodies.
Use beta Hydroxybutyrate (beta HB) Assay Kit
ab83390
to measure the beta hydroxybutyrate element of ketone bodies.
To measure both the beta hydroxybutyrate and acetoacetate elements of ketone bodies in a single assay kit, use Ketone Body Assay Kit
ab318308
Other notes
This product was previously called K650 Biovision Acetoacetate Colorimetric Assay Kit. Biovision was acquired by Abcam in 2021.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C
Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Acetoacetate is a four-carbon ketone body that plays a central role in energy metabolism particularly during periods of fasting or carbohydrate restriction. It has a molecular mass of 102.09 g/mol. Acetoacetate is mainly produced in the liver's mitochondria and can be converted to either acetone or beta-hydroxybutyrate another key ketone body dependent on the enzymatic conditions present. This conversion process involves several enzymes including acetoacetate decarboxylase for acetoacetate to acetone and beta-hydroxybutyrate dehydrogenase for acetoacetate to beta-hydroxybutyrate.
Biological function summary
The presence of acetoacetate serves as an alternative energy source for many tissues including the brain heart and muscles when glucose availability is low. Acetoacetate and its conversion to beta-hydroxybutyrate are important during states of low-carbohydrate intake prolonged exercise or starvation. These compounds bypass the blood-brain barrier to supply energy to neurons. Acetoacetate itself does not form part of a larger protein complex but interacts directly with enzymes to mediate its functions.
Pathways
Acetoacetate is predominantly involved in the ketogenic pathway where it is synthesized from the breakdown of fatty acids. This pathway plays an important role in maintaining energy balance during carbohydrate scarcity. In the citric acid cycle acetoacetate is linked to acetyl-CoA aiding in the generation of ATP. Proteins like succinyl-CoA:3-ketoacid CoA transferase facilitate acetoacetate's conversion into energy-yielding substrates interrelating with both pathways for effective energy modulation.
Abnormal levels of acetoacetate can relate to conditions such as diabetic ketoacidosis and alcoholic ketoacidosis. In diabetic ketoacidosis elevated acetoacetate levels are due to insulin deficiency leading to excessive ketone body production often accompanied by high beta-hydroxybutyrate levels. This condition links acetoacetate to insulin a critical regulatory hormone. In alcoholic ketoacidosis chronic alcohol consumption results in acetoacetate accumulation due to disrupted carbohydrate metabolism highlighting its role in metabolic imbalances.
