Acetoacetate Assay Kit (Colorimetric) ab180875 adapts the traditional qualitative assay using dipsticks, and sodium nitroferricyanide as a chromophore, with a modification that provides a sensitive method to quantitate endogenous levels of AcAc in human blood and urine.
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- Nanomaterials (Basel, Switzerland) 12:2022Punicalagin and Ketogenic Amino Acids Loaded Organic Lipid Carriers Enhance the Bioavailability, Mitochondrial β-Oxidation, and Ketogenesis in Maturing Adipocytes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPandurangan Subash-Babu,Nouf Al-Numair,Tahani Almuzaini,Jegan Athinarayanan,Ali Abdullah AlshatwiPubMed 35159714
- FEBS open bio 11:1893-19062021The role of SAMM50 in non-alcoholic fatty liver disease: from genetics to mechanisms.Applications:Unspecified applicationReactive species:Unspecified reactive speciesZuyin Li,Weixing Shen,Gang Wu,Changjiang Qin,Yijie Zhang,Yupeng Wang,Guohe Song,Chao Xiao,Xin Zhang,Guilong Deng,Ruitao Wang,Xiaoliang WangPubMed 33728819
- Journal of the American Heart Association 9:e0130392020Increased Glucose Availability Attenuates Myocardial Ketone Body Utilization.Applications:Unspecified applicationReactive species:Unspecified reactive speciesManoja K Brahma et. al.PubMed 32750298
- GeroScience 42:667-6862020Short-term time-restricted feeding is safe and feasible in non-obese healthy midlife and older adults.Applications:Unspecified applicationReactive species:Unspecified reactive speciesChristopher R Martens et. al.PubMed 31975053
- Diagnostics (Basel, Switzerland) 9:2019Blood Ketone Bodies and Breath Acetone Analysis and Their Correlations in Type 2 Diabetes Mellitus.Applications:Unspecified applicationReactive species:Unspecified reactive speciesValentine Saasa et. al.PubMed 31861135
- Stroke 49:2173-21812018Imaging the Effects of β-Hydroxybutyrate on Peri-Infarct Neurovascular Function and Metabolism.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPaolo Bazzigaluppi,Evelyn M Lake,Tina L Beckett,Margaret M Koletar,Iliya Weisspapir,Stefan Heinen,James Mester,Aaron Lai,Rafal Janik,Adrienne Dorr,JoAnne McLaurin,Greg J Stanisz,Peter L Carlen,Bojana StefanovicPubMed 30354983
- Translational psychiatry 8:662018Prebiotic attenuation of olanzapine-induced weight gain in rats: analysis of central and peripheral biomarkers and gut microbiota.Applications:FuncSReactive species:Unspecified reactive speciesAmy Chia-Ching Kao et. al.PubMed 29540664
- Frontiers in physiology 8:8062017Ketone Diester Ingestion Impairs Time-Trial Performance in Professional Cyclists.Applications:Unspecified applicationReactive species:Unspecified reactive speciesJill J Leckey et. al.PubMed 29109686
- Oncotarget 8:78614-786322017Identification of LRP-1 as an endocytosis and recycling receptor for β1-integrin in thyroid cancer cells.
- PloS one 12:e01744772017Ketogenic diet improves the spatial memory impairment caused by exposure to hypobaric hypoxia through increased acetylation of histones in rats.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMing Zhao et. al.PubMed 28355243
Key facts
- Detection method
- Colorimetric
- Sample types
- Urine, Plasma, Other biological fluids
- Assay type
- Quantitative
- Reactive species
- Mammals
- Assay time
- 25m
- Sensitivity
- < 25 µM
Target data
What's included?
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Acetoacetate Assay Kit (Colorimetric) ab180875 adapts the traditional qualitative assay using dipsticks, and sodium nitroferricyanide as a chromophore, with a modification that provides a sensitive method to quantitate endogenous levels of AcAc in human blood and urine.
Key facts
- Detection method
- Colorimetric
- Sample types
- Urine, Plasma, Other biological fluids
- Assay type
- Quantitative
- Reactive species
- Mammals
- Assay time
- 25m
- Assay Platform
- Microplate reader
- Sensitivity
- < 25 µM
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- -20°C
- Appropriate long-term storage conditions
- -20°C
- Storage information
- -20°C
Notes
Acetoacetate Assay Kit (Colorimetric) ab180875 adapts the traditional qualitative assay using dipsticks, and sodium nitroferricyanide as a chromophore, with a modification that provides a sensitive method to quantitate endogenous levels of AcAc in human urine.
In this non-enzymatic assay, AcAc reacts with a substrate 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
This product is manufactured by BioVision, an Abcam company and was previously called K650 Acetoacetate Colorimetric Assay Kit. K650-100 is the same size as the 100 test size of ab180875.
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 ((CH3)2CO). Ketone bodies [βHB: 78%; AcAc: 20% & (CH3)2CO: 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.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
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.
- Associated diseases and disorders
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.
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5 product images
Functional Studies - Acetoacetate Assay Kit (Colorimetric) (ab180875)
Functional assay - 180875 Acetoacetate Assay Kit (Colorimetric).
Acetoacetate measured in urine samples showing concentration (micromolar).
Functional Studies - Acetoacetate Assay Kit (Colorimetric) (ab180875)
Functional assay - 180875 Acetoacetate Assay Kit (Colorimetric).
Colorimetric standard curve: mean of duplicates (+/-SD) with background readings subtracted.
Functional Studies - Acetoacetate Assay Kit (Colorimetric) (ab180875)
Measurement of Acetoacetate in urine.
Samples were deproteinized by spin filtering. 100 μL of undiluted urine was spiked with known amounts of AcAc (30 nmol).
Assays were performed following the kit protocol.
Functional Studies - Acetoacetate Assay Kit (Colorimetric) (ab180875)
Standard Curve for equimolar amounts of Acetoacetate, β-HB and Acetone (4°C, 100 min.). The other two ketone bodies β-hydroxybutyrate or Acetone are not detected in the assay due to much lower sensitivity (β-HB) and much lower concentration (Acetone).
Functional Studies - Acetoacetate Assay Kit (Colorimetric) (ab180875)
Time course experiment.
Time course of color development at 4°C and 25°C (100 nmol Acetoacetate).
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