Product datasheet
Acetoacetate Assay Kit (Colorimetric) ab180875
5 Images
Overview
Product name
Acetoacetate Assay Kit (Colorimetric)
Detection method
Colorimetric
Sample type
Urine, Serum, Plasma, Other biological fluids, Whole Blood
Assay type
Quantitative
Sensitivity
< 25 µM
Assay duration
Multiple steps standard assay
Species reactivity
Reacts with: Human
Predicted to work with: all Mammals
Product overview
Acetoacetate (AcAc), a ß-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-ß-hydroxybutyrate (ß-HB), or decarboxylated producing acetone ((CH3)2CO).
Ketone bodies [ßHB: 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.
Abcam’s Acetoacetate Assay Kit (Colorimetric) (ab180875) has adapted that principle 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-ß-hydroxybutyrate. The assay kit can detect samples containing acetoacetate as low as 25
µM.
Tested applications
Functional Studies
Platform
Microplate
Properties
Storage instructions
Store at -20°C. Please refer to protocols.
Components
Identifier
Acetoacetate Assay Buffer
100 tests
1 x 15ml
Acetoacetate Standard (Lyophilized)
Yellow
1 vial
Acetoacetate Substrate
Amber
1 x 1ml
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Applications
Our Abpromise guarantee covers the use of ab180875 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application
Abreviews
Functional Studies
Notes
Use at an assay dependent concentration.
Acetoacetate Assay Kit (Colorimetric) images
Acetoacetate measured in urine samples
showing concentration (micromolar).
Functional assay - 180875 Acetoacetate Assay
Kit (Colorimetric)
Colorimetric standard curve: mean of
duplicates (+/-SD) with background readings
subtracted.
Functional assay - 180875 Acetoacetate Assay
Kit (Colorimetric)
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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.
Measurement of Acetoacetate in urine
Standard Curve for equimolar amounts of
Acetoacetate, β-HB and Acetone (4°C, 100
min.). The other two ketone bodies βhydroxybutyrate or Acetone are not detected
in the assay due to much lower sensitivity (βHB) and much lower concentration (Acetone).
Standard Curve
Time course of color development at 4°C and
25°C (100 nmol Acetoacetate).
Time course experiment
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Replacement or refund for products not performing as stated on the datasheet
Valid for 12 months from date of delivery
Response to your inquiry within 24 hours
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If the product does not perform as described on this datasheet, we will offer a refund or replacement. For full details of the Abpromise,
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Terms and conditions
Guarantee only valid for products bought direct from Abcam or one of our authorized distributors
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