Acetoacetate Colorimetric Assay Kit (BN00874)

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BN00874
  • Acetoacetate Colorimetric Assay Kit (BN00874)
  • Acetoacetate Colorimetric Assay Kit (BN00874)
  • Acetoacetate Colorimetric Assay Kit (BN00874)
€876

Description

ELISA Kit Technical ManualMSDS

Acetoacetate Colorimetric Assay Kit

Acetoacetate (AcAc), a Beta-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-Beta-hydroxybutyrate (Beta-HB), or decarboxylated producing acetone (CH3)2CO). Ketone bodies (BetaHB: 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. Assay Genie's Acetoacetate Assay Kit 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-Beta-hydroxybutyrate. The assay kit can detect samples containing acetoacetate as low as 25 µM.

Figure: a) Time course of color development at 4°C and 25°C (100 nmol Acetoacetate). b) 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). c) 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.

Key Information Description

Product SKU

BN00874

Size

100 assays

Detection Method

Absorbance (550 nm)

Applications

  • Measurement of acetoacetate in various biological samples.
  • Analysis of acetoacetate in pathological conditions such as Type I diabetes, alcoholism, etc.

Features and Benefits

  • Specific for acetoacetate and does not detect 3-Beta-hydroxybutyrate
  • The assay can detect less than 25 µM of acetoacetate in various sample types.

Kit Components

  • Acetoacetate Assay Buffer
  • Acetoacetate Standard (Lyophilized)
  • Acetoacetate Substrate

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

USAGE

For Research Use Only! Not For Use in Humans.

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