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

Amplite® Fluorimetric Beta-Hydroxybutyrate (Ketone Body) Assay Kit is a fluorimetric assay kit for quantification of beta-hydroxybutyrate in biological samples.

Detection Principle: Uses an enzyme-coupled reaction to generate NADH.
Sensitivity: Detects beta-HB down to 1.4 µM.
Detection Chemistry: Uses a fluorescent NADH sensor.
Instrument Compatibility: Fluorescence detection at Ex/Em 571/585 nm with a microplate reader.

&beta;-Hydroxybutyrate (&beta;-HB) dose response was measured with Amplite&trade; Fluorimetric &beta;-Hydroxybutyrate Assay Kit on a solid black 96-well plate using a Gemini microplate reader.

Protocol

SDS

Catalog	Size	Price	Quantity
13831	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, while acetone is the third abundant ketone body. Normally these two predominant ketone bodies are present in small amounts in the blood during fasting 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® Fluorimetric Beta-Hydroxybutyrate Assay Kit offers a sensitive fluorescent assay for measuring Beta-HB levels in biological samples. This assay is based on an enzyme coupled reaction of Beta-HB, in which the product NADH can be specifically monitored by a fluorescent NADH sensor. The fluorescence signal can be measured by a fluorescence microplate reader. With this Fluorimetric Beta-hydroxybutyrate Assay Kit, we were able to detect as low as 1.4 µM Beta-HB in a 100 µL reaction volume.

Citations

View all 4 citations: Citation Explorer

Pin1 mediates metabolic dysfunction-associated steatohepatitis in mice fed high-fat, high-cholesterol diet by regulating both PPAR$\alpha$ and acetyl CoA carboxylase

Authors:

Nakatsu, Yusuke and Sano, Tomomi and Nakanishi, Mikako and Matsunaga, Yasuka and Kanna, Machi and Kanematsu, Takashi and Asano, Tomoichiro

Journal:

Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease (2025): 168087

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

Highly differentiated human fetal RPE cultures are resistant to the accumulation and toxicity of lipofuscin-like material

Authors:

Zhang, Qitao and Presswalla, Feriel and Calton, Melissa and Charniga, Carol and Stern, Jeffrey and Temple, Sally and Vollrath, Douglas and Zacks, David N and Ali, Robin R and Thompson, Debra A and others,

Journal:

Investigative ophthalmology \& visual science (2019): 3468--3479

A platform for assessing outer segment fate in primary human fetal RPE cultures

Authors:

Zhang, Qitao and Presswalla, Feriel and Feathers, Kecia and Cao, Xu and Hughes, Bret A and Zacks, David N and Thompson, Debra A and Miller, Jason ML

Journal:

Experimental eye research (2018)

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

Fluorescence microplate reader
Excitation	540 nm
Emission	590 nm
Cutoff	570 nm
Recommended plate	Solid black

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Page updated on October 8, 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)