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BHB-Glo™ (Ketone Body) Assay Technical Manual

Instructions for Use of Product(s)
JE9500, JE9600

Literature # TM713

The BHB-Glo™ (Ketone Body) Assay is a bioluminescent assay for rapid, selective, sensitive detection of β-hydroxybutyratem (BHB) in biological samples. β-hydroxybutyrate is a ketone body that is synthesized in the liver from fatty acids during low glucose conditions, such as starvation or prolonged exercise. BHB acts as an energy source for peripheral tissues and is believed to have signaling properties that involve many cellular functions.
The BHB-Glo™ (Ketone Body) Assay couples BHB oxidation and NADH production with a bioluminescent NADH detection system. When BHB detection reagent is added to a sample at a 1:1 ratio, the coupled-enzyme reactions are initiated and run simultaneously. The luminescent signal is proportional to the amount of BHB in the sample and increases until all BHB is consumed, at which point a stable luminescent signal is achieved.

Summary of Changes
The following changes were made to the 1/24 revision of this document:
1. In the legend for Figure 7, 2μM BHB standard was changed to 4μM BHB standard. Also in this figure legend, 6μM HCl was changed to 6N HCl.

Revised 1/24.