EVALUATION OF LUMIGEN HYPERBLU TECHNOLOGY FOR THE HIGH THROUGHPUT DETECTION OF HYDROGEN PEROXIDE
David Schumm, Robert A. Eickholt Beckman Coulter Inc., Southfield, MI
Jessica L Schneck, Elsie Diaz GlaxoSmithKline, Collegeville, PA
HyPerBlu technology is a uniquely capable reagent for direct detection of hydrogen peroxide over a range from 25 nmol to 500 fmol. Enzymatic production of hydrogen peroxide has become an attractive target for high-throughput screening research. Traditional resorufin based fluorescent tools such as Amplex RedTM (N-acetyl-3,7-dihydroxyphenoxazine) suffer from a more narrow dynamic range, and are limited in use by their stability. The chemistry of the HyPerBlu formulation does not require HRP or other biological intermediary. The active compound is a stable dioxetane which produces a strong chemiluminescent signal upon reaction with hydrogen peroxide. HyPerBlu substrate is stable for over a month at room temperature, and over one year at 2-8 oC . HyPerBlu reagent can maintain its overall intensity, as well as linear response and sensitivity, over an extended period of time. This stability, combined with its robust compatibility with common assay additives (e.g., DMSO), make Lumigen HyPerBlu an enabling new technology for use in HTS assays. An assay developed to study xanthine oxidase was chosen to evaluate this detection technology.
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