Hydrogen Peroxide is produced in biological systems by controlled pathways at low concentrations that impact on cell signalling. At higher concentrations inflammatory cells produce local intense amounts of this oxidant to kill pathogens. In the development of human disease uncontrolled formation of hydrogen peroxide from the mitochondrial respiratory chain and enzymes such as xanthine oxidase can occur (Prof. Victor Darley-Usmar, University of Alabama, personal communication). Hydrogen peroxide concentrations at µM level elicit a broad spectrum of cellular responses ranging from mitogenic growth stimulation to apoptosis and necrosis. PeroxiLuminol is a hydrogen peroxide chemiluminescence bioassay kit developed by WPI to provide an accurate analytical tool by which to measure hydrogen peroxide formation in the living system. The kit is designed to complement WPI’s new range of hydrogen peroxide electrochemical sensors. When combined together these detection methods will greatly aid hydrogen peroxide research.

Assay Principle: Horseradish peroxidase (HRP) reacts with hydrogen peroxide to form reactive Compound I and Compound II, which in turn oxidize luminol with concomitant production of chemiluminescence (see reaction scheme below). The PeroxiLuminol Kit has been optimized so that hydrogen peroxide concentration is linearly proportional to luminescence intensity. Our formulation assures that luminescence intensity doesn’t decay too quickly so that measurement can be made accurately and precisely. The slow decay rate also allows luminescence measurement with simple luminometers that may not include injection/mixing devices. The figure above shows hydrogen peroxide standards as assayed by the HRP-catalyzed luminescence method.

Benefits

• Determine hydrogen peroxide concentration in minutes.
• Simple and extensively tested method with an increased sensitivity.
• More stable signals.
• Inexpensive measurement instrument.

Note: This kit is designed to assay hydrogen peroxide in cell cultures. Samples with high haemoglobin content and/or metal content may not be suitable for assay. Due to continued development, the kit format and performance parameters may be changed without a notice. Contact WPI for further information.

This kit provides material for 100 assays in cuvettes or test tubes (assay volume: 1 mL/assay).
 

H. P. Misra & P. M. Squatrito (1982), Arch. Biochem. Biophys. 215, 59.
G. H. G. Thorpe & L. J. Kricka (1986), Methods in Enzymology 133, 331.