Acetoacetate Assay Kit (Colorimetric) (ab180875)
Key features and details
- Assay type: Quantitative
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 25 min
- Sample type: Other biological fluids, Plasma, Urine, Whole Blood
- Sensitivity: 25 µM
Overview
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Product name
Acetoacetate Assay Kit (Colorimetric) -
Detection method
Colorimetric -
Sample type
Urine, Plasma, Other biological fluids, Whole Blood -
Assay type
Quantitative -
Sensitivity
< 25 µM -
Assay time
0h 25m -
Species reactivity
Reacts with: Mammals, Other species -
Product overview
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.
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 -
Notes
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.
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Platform
Microplate reader
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests Acetoacetate Assay Buffer 1 x 15ml Acetoacetate Standard 1 vial Acetoacetate Substrate 1 x 1ml
Associated products
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Related Products
Images
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Acetoacetate measured in urine samples showing concentration (micromolar).
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Colorimetric standard curve: mean of duplicates (+/-SD) with background readings subtracted.
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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.
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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).
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Time course of color development at 4°C and 25°C (100 nmol Acetoacetate).
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab180875 has been referenced in 10 publications.
- Subash-Babu P et al. Punicalagin and Ketogenic Amino Acids Loaded Organic Lipid Carriers Enhance the Bioavailability, Mitochondrial ß-Oxidation, and Ketogenesis in Maturing Adipocytes. Nanomaterials (Basel) 12:N/A (2022). PubMed: 35159714
- Li Z et al. The role of SAMM50 in non-alcoholic fatty liver disease: from genetics to mechanisms. FEBS Open Bio 11:1893-1906 (2021). PubMed: 33728819
- Martens CR et al. Short-term time-restricted feeding is safe and feasible in non-obese healthy midlife and older adults. Geroscience 42:667-686 (2020). PubMed: 31975053
- Brahma MK et al. Increased Glucose Availability Attenuates Myocardial Ketone Body Utilization. J Am Heart Assoc 9:e013039 (2020). PubMed: 32750298
- Saasa V et al. Blood Ketone Bodies and Breath Acetone Analysis and Their Correlations in Type 2 Diabetes Mellitus. Diagnostics (Basel) 9:N/A (2019). PubMed: 31861135
- Kao AC et al. Prebiotic attenuation of olanzapine-induced weight gain in rats: analysis of central and peripheral biomarkers and gut microbiota. Transl Psychiatry 8:66 (2018). Functional Studies . PubMed: 29540664
- Bazzigaluppi P et al. Imaging the Effects of ß-Hydroxybutyrate on Peri-Infarct Neurovascular Function and Metabolism. Stroke 49:2173-2181 (2018). PubMed: 30354983
- Zhao M et al. Ketogenic diet improves the spatial memory impairment caused by exposure to hypobaric hypoxia through increased acetylation of histones in rats. PLoS One 12:e0174477 (2017). PubMed: 28355243
- Leckey JJ et al. Ketone Diester Ingestion Impairs Time-Trial Performance in Professional Cyclists. Front Physiol 8:806 (2017). PubMed: 29109686
- Theret L et al. Identification of LRP-1 as an endocytosis and recycling receptor for ß1-integrin in thyroid cancer cells. Oncotarget 8:78614-78632 (2017). WB ; Human . PubMed: 29108253