Milk, Urine, Serum, Plasma, Other biological fluids
< 4 µM
Multiple steps standard assay
Abcam's β-Hydroxybutyrate Assay Kit (Fluorometric) (ab180876) offers simplicity, enhanced sensitivity, and can be adapted to high-throughput applications. The assay is based on enzymatic oxidation of β-HB that results in generation of fluorescent signal (Ex/Em = 535/587 nm) which is directly proportional to the amount of β-HB. The kit offers an excellent alternative for measurement of β-HB when biological sample quantities are limited or subketogenic levels are suspected. The assay can detect β-Hydroxybutyrate as low as 4 µM in a variety of biological samples.
β-Hydroxybutyrate (β-HB) is the most abundant of the ketone bodies (~78% of total ketone bodies in blood). Ketoacidosis (high concentrations of the ketone bodies) can be caused by a variety of conditions, such as diabetes, alcoholism and severe starvation. Ketoacidosis can be fatal if not treated. Additionally, ketogenic diets can increase the concentration of ketone bodies slightly in healthy individuals.
Diabetic ketoacidosis occurs when circulating insulin levels drop to very low levels, shutting off the supply of glucose to the body. The physiological response is for the liver to produce ketone bodies (acetoacetate, acetone, and primarily beta hydroxybutyrate) from the acetyl CoA produced from fatty acid oxidation. The very high rate of ketone body production outstrips the body's ability to utilize them as an energy source and the blood concentration builds up. As rather strong acids, they result in a significant drop in blood pH.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
ß-Hydroxybutyrate Standard Curve
Measurement of ß-Hydroxybutyrate concentration
Measurement of β-Hydroxybutyrate concentration in human urine (2.5 µL), serum (2.5 µL) & 2% Milk (5 µL). All samples were deproteinized using 10 kDa Spin Column & spiked with known amount of β-Hydroxybutyrate (400 pmol). Assays were performed following the kit protocol.