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Implementation of a Continuous, Enzyme-Coupled Fluorescence Assay for High-Throughput Analysis of Glutamate-Producing Enzymes

โœ Scribed by Kathryn E. McElroy; Peter J. Bouchard; Mark R. Harpel; Kurumi Y. Horiuchi; Kelley C. Rogers; Dennis J. Murphy; Thomas D.Y. Chung; Robert A. Copeland

Book ID
102564713
Publisher
Elsevier Science
Year
2000
Tongue
English
Weight
77 KB
Volume
284
Category
Article
ISSN
0003-2697

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โœฆ Synopsis

Enzymatic formation of glutamate is critical to numerous biological pathways. However, current methods for assaying the activities of glutamate-forming enzymes are not particularly suitable for highthroughput screening in drug discovery. We present a continuous-read, fluorometric assay for high-throughput analysis of glutaminases. This assay is adapted to a microplate format and employs glutamate oxidase and horseradish peroxidase to couple glutamate formation to production of the fluorescent reporter molecule, resorufin, for enhancement of sensitivity (M.

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