Amplite® Colorimetric Beta-Hydroxybutyrate (Ketone Body) Assay Kit

Amplite® Colorimetric Beta-Hydroxybutyrate (Ketone Body) Assay Kit is a colorimetric assay kit for quantifying beta-hydroxybutyrate (beta-HB) in biological samples through an enzyme-coupled NADH readout.

Colorimetric detection: Measures NADH generated by an enzyme-coupled beta-HB reaction using an NADH sensor.
High sensitivity: Detects beta-HB concentrations as low as 4 µM in a 100 µL reaction volume.
Enzyme-coupled chemistry: Converts beta-HB into a proportional NADH-dependent colorimetric signal.
Microplate compatible: Read absorbance at 570/610 nm using a clear-bottom plate.
Applications: Suitable for beta-HB quantitation in biological samples and ketone metabolism studies.

&beta;-Hydroxybutyrate (&beta;-HB) dose response was measured with Amplite&trade; Colorimetric &beta;-Hydroxybutyrate Assay Kit on a black wall/clear bottom 96-well plate using a SpectraMax microplate reader (Molecular Devices).

Protocol

SDS

Catalog	Size	Price	Quantity
13830	200 Tests	Price

Overview

Ketone bodies are produced by the liver and used peripherally as an energy source when blood glucose levels drop. The two main ketone bodies are beta-hydroxybutyrate (beta-HB) and acetoacetate (AcAc), while acetone is the third abundant ketone body. Normally these two predominant ketone bodies are present in small amounts in the blood during fasting (low food intake) and prolonged exercise. In patients who have diabetes, alcohol or salicylate poisoning, hormone deficiency, childhood hypoglycemia and other acute disease states, large quantities of ketone bodies are found in the blood. The over-production and accumulation of ketone bodies in the blood (ketosis) can lead to pathological metabolic acidosis (ketoacidosis). In extreme cases, ketoacidosis can be fatal. Blood ketone testing methods that quantify beta-HB, the predominant ketone body in the blood (approximately 75%) have been used for diagnosing and monitoring treatment of ketoacidosis. Amplite® Colorimetric Beta-Hydroxybutyrate Assay Kit offers a sensitive colorimetric assay for measuring beta-HB levels in biological samples. This assay is based on an enzyme coupled reaction of beta-HB, in which the NADH product can be specifically monitored by a NADH sensor. The signal can be measured by an absorbance microplate reader with the OD ratio at the wavelength of 570 nm to 610 nm. With this Amplite® Colorimetric Beta-Hydroxybutyrate Assay Kit, we were able to detect as low as 4 µM beta-HB in a 100 µL reaction volume.

Citations

View all 5 citations: Citation Explorer

Reassessing the accuracy of enzyme-based assays for $\beta$-hydroxybutyrate production by the retinal pigment epithelium

Authors:

Autterson, Gillian and Han, John Yeong Se and Philp, Nancy and Miller, Jason Matthew Lewis

Journal:

Investigative Ophthalmology \& Visual Science (2023): 4466--4466

A combination of ketogenic diet and voluntary exercise ameliorates anxiety and depression-like behaviors in Balb/c mice

Authors:

Gumus, Hikmet and Ilgin, Rabia and Koc, Basar and Yuksel, Oguz and Kizildag, Servet and Guvendi, Guven and Karakilic, Asli and Kandis, Sevim and Hosgorler, Ferda and Ates, Mehmet and others,

Journal:

Neuroscience Letters (2022): 136443

$\beta$-hydroxybutyrate suppresses NLRP3 inflammasome-mediated placental inflammation and lipopolysaccharide-induced fetal absorption

Authors:

Hirata, Yoshiki and Shimazaki, Sayaka and Suzuki, Sae and Henmi, Yuka and Komiyama, Hiromu and Kuwayama, Takehito and Iwata, Hisataka and Karasawa, Tadayoshi and Takahashi, Masafumi and Takahashi, Hironori and others,

Journal:

Journal of Reproductive Immunology (2021): 103433

Stopover ecology of moult migrant Swainson's thrushes {\guillemotleft}Catharus ustulatus{\guillemotright}

Authors:

Morales, Ana

Point-of-care blood analyzers measure the nutritional state of eighteen free-living bird species

Authors:

Morales, Ana and Frei, Barbara and Leung, Casey and Titman, Rodger and Whelan, Shannon and Benowitz-Fredericks, Z Morgan and Elliott, Kyle H

Journal:

Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology (2020): 110594

References

View all 54 references: Citation Explorer

Surface glycosylation of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) membrane for selective adsorption of low-density lipoprotein

Authors:

Wang W, Lan P.

Journal:

J Biomater Sci Polym Ed (2014): 2094

Biocompatibility of electrospun poly(3-hydroxybutyrate) and its composites scaffoldsfor tissue engineering.

Authors:

Zharkova, II, Staroverova OV, Voinova VV, Andreeva NV, Shushckevich AM, Sklyanchuk ED, Kuzmicheva GM, Bespalova AE, Akulina EA, Shaitan KV, Olkhov AA.

Journal:

Biomed Khim (2014): 553

Gamma-hydroxybutyrate (GHB) induces cognitive deficits and affects GABAB receptors and IGF-1 receptors in male rats

Authors:

Johansson J, Gronbladh A, Hallberg M.

Journal:

Behav Brain Res (2014): 164

Protective effects of exogenous beta-hydroxybutyrate on paraquat toxicity in rat kidney

Authors:

Wei T, Tian W, Liu F, Xie G.

Journal:

Biochem Biophys Res Commun (2014): 666

Lotus-leaf-like topography predominates over adsorbed ECM proteins in poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) surface/cell interactions

Authors:

Zheng J, Li D, Yuan L, Liu X, Chen H.

Journal:

ACS Appl Mater Interfaces (2013): 5882

Usage and storage

Intended use

Research Use Only (RUO)

Instrument settings

Absorbance microplate reader
Absorbance	570/610 nm
Recommended plate	Clear bottom

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Page updated on October 1, 2026

Component A: Enzyme Mix	2 bottles (lyophilized powder)
Component B: Assay Buffer	1 bottle (10 mL)
Component C: NAD	1 vial
Component D: β-Hydroxybutyrate (β-HB) Standard	1 vial (10 µL)