EnzyChrom™ Ketone Body Assay Kit
Protocol SDS  Printed Protocols and SDSs are not provided with kits.

Application
For quantitative determination of ketone body (acetoacetate and 3-hydroxybutyrate) and evaluation of drug effects on its metabolism.

Key Features
Sensitive and accurate. Uses 10 μL sample. Linear detection range of 0.12 to 8 mM for each ketone body in 96-well plate assay.

Convenient. The procedure involves adding a single working reagent, and reading the optical density at room temperature.

High-throughput. Can be automated as a high-throughput 96-well plate assay for many samples per day.

Method
OD340nm

Samples
Serum, plasma, urine etc

Species
All

Procedure
30 min

Size
200 tests

Detection Limit
0.12 mM

Shelf Life
6 months

More Details
Ketone bodies (acetoacetic acid and 3-hydroxybutyric acid) are produced in the liver mainly from oxidation of fatty acids, and are normally present at low concentrations in urine and blood. Increased ketone concentrations in the blood may lead to metabolic acidosis, which has been associated with diabetes, childhood hypo-glycaemia, growth hormone deficiency, alcohol or salicylate intoxication and inborn errors of metabolism. Simple, direct and automation-ready procedures for measuring acetoacetic acid (AcAc) and 3-hydroxybutyric acid (BOH) are very desirable. BioAssay Systems EnzyChrom™ ketone body assay is based on 3-hydroxybutyrate dehydrogenase catalyzed reactions, in which the change in NADH absorbance, measured at 340nm, is directly related to the AcAc and BOH concentrations,

1. Can serum and plasma samples be stored frozen?



β-hydroxybutyrate is stable and can be detected in samples that had been stored frozen for more than a month. Acetoacetate (AcAc), on the other hand, is an unstable compound which becomes rapidly decarboxylated. We recommend to use fresh serum or freeze serum samples at -80°C for up to one month.

2. I seem to be getting very low levels of BOH and AcAc in freshly drawn blood. Perhaps this is because they are breaking down within the hour it takes me to process the plasma out.



In healthy individuals, normal blood values for ketone bodies are actually very low or not detectable. Elevated blood levels of ketone bodies (ketosis) can be caused for example by diabetes, alcohol intoxication, starvation, or physical exercise. You can check for the stability of your analyte by setting aside an aliquot of your sample and add the standard as a positive control.

For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions
Cui, A., Fan, H., Zhang, Y., Zhang, Y., Niu, D., Liu, S. & Liu, Y. (2019). Dexamethasone-induced Kruppel-like factor 9 expression promotes hepatic gluconeogenesis and hyperglycemia. The Journal of clinical investigation 130:2266-2278. Assay: Ketone body in mice serum.

Saito, T., Kuma, A., Sugiura, Y., Ichimura, Y., Obata, M., Kitamura, H. & Saito, I. (2019). Autophagy regulates lipid metabolism through selective turnover of NCoR1. Nature communications 10(1):1567. Assay: Ketone body in mice plasma.

Herrick, K. J., Hippen, A. R., Kalscheur, K. F., Schingoethe, D. J., Ranathunga, S. D., Anderson, J. L. & Van Eys, J. E. (2018). Infusion of butyrate affects plasma glucose, butyrate, and beta-hydroxybutyrate but not plasma insulin in lactating dairy cows. Journal of dairy science, 101(4), 3524-3536. Assay: Ketone body in cow plasma.

Iso, T., Haruyama, H., Sunaga, H., Matsui, H., Matsui, M., Tanaka, R. & Hanaoka, H. (2018). CD36 is indispensable for nutrient homeostasis and endurance exercise capacity during prolonged fasting. Physiological reports, 6(19), e13884. Assay: Ketone body in mice serum.

Lyons, K. (2018). Physiological consequences of environmental contamination in an elasmobranch with matrotrophic histotrophy, the Round Stingray (Urobatis halleri). Assay: Ketone body in stingrays plasma.

Bisgaier, Charles L., and Daniela Carmen Oniciu (2017). Treatment of NASH with gemcabene. U.S. Patent No. 9,849,104 Assay: Ketone body in mice plasma.

Goossens, C., Marques, M. B., Derde, S., Vander Perre, S., Dufour, T., Thiessen, S. E. & De Bock, K. (2017). Premorbid obesity, but not nutrition, prevents critical illness-induced muscle wasting and weakness. Journal of cachexia, sarcopenia and muscle, 8(1), 89-101. Assay: Ketone body in mice serum.

Herrick, K. J., Hippen, A. R., Kalscheur, K. F., Schingoethe, D. J., Casper, D. P., Moreland, S. C., & Van Eys, J. E. (2017). Single-dose infusion of sodium butyrate, but not lactose, increases plasma beta-hydroxybutyrate and insulin in lactating dairy cows. Journal of dairy science, 100(1), 757-768. Assay: Ketone body in cow plasma.

Driver, C. (2016). Metformin does not prevent glucose intolerance but improves renal function and reduces oxidative stress in type 1 diabetes (Doctoral dissertation). Assay: Ketone body in rat serum and urine.

Murunga, A. N., Miruka, D. O., Driver, C., Nkomo, F. S., Cobongela, S. Z., & Owira, P. M. (2016). Grapefruit derived flavonoid naringin improves ketoacidosis and lipid peroxidation in type 1 diabetes rat model. PloS one, 11(4), e0153241. Assay: Ketone body in rat serum and urine.

Neondo, M. A. (2016). The effects of naringin on glucose tolerance and ketoacidosis in type 1 diabetes (Doctoral dissertation). Assay: Ketone body in rat serum and urine.

Syamsunarno, MRAA et al (2014). Fatty Acid Binding Protein 4 and 5 Play a Crucial Role in Thermogenesis under the Conditions of Fasting and Cold Stress. PloS one 9.3: e90825. Assay: Ketone body in mouse serum.

Fujieda, Y et al (2013). Inflammation and Resolution Are Associated with Upregulation of Fatty Acid beta-Oxidation in Zymosan-Induced Peritonitis. PloS one 8.6: e66270. Assay: Ketone body in mouse liver tissue.

Martin-Murphy, BV et al (2013). Increased susceptibility of natural killer T-cell-deficient mice to acetaminophen-induced liver injury. Hepatology 57.4: 1575-1584. Assay: Ketone body in mouse serum.

Syamsunarno, MRAA et al (2013). A Critical Role of Fatty Acid Binding Protein 4 and 5 (FABP4/5) in the Systemic Response to Fasting. PloS one 8.11: e79386. Assay: Ketone body in mouse serum.

Qi S et al (2012). Comparison of the metabolic profiling of hepatitis B virus-infected cirrhosis and alcoholic cirrhosis patients by using (1) H NMR-based metabonomics. Hepatol Res 42(7):677-85. Assay: Ketone body in human serum.

Klusonova P et al (2011) Local metabolism of glucocorticoids in Prague hereditary hypertriglyceridemic rats--effect of hypertriglyceridemia and gender. Steroids 76(12):1252-9. Assay: Ketone Body in rat Hepatocyte culture.

Sharma NS et al (2011). Metabolic profiling based quantitative evaluation of hepatocellular metabolism in presence of adipocyte derived extracellular matrix. PLoS One 6(5):e20137. Assay: Ketone body in rat hepatocyte culture.

Sharma NS et al (2011). Metabolic profiling based quantitative evaluation of hepatocellular metabolism in presence of adipocyte derived extracellular matrix. PLoS One 6(5):e20137. Assay: Ketone Body in human serum.

To find more recent publications, please click here.
Please inquire or request assay service or call 1-510-782-9988 x 2.
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EnzyChrom™ Ketone Body Assay Kit
Catalog No: EKBD-100
Price: $389    Qty:
For orders of 10 or more kits, please call 1-510-7829988x1 or email us for best pricing and/or bulk order.

Shipping: On Ice
Shipment: Fedex Service
Delivery: 1-2 days (US), 3-6 days (Intl) Storage: -20°C

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