Stable reagent and kinetic assay for alpha-amylase

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase

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435 4, 435 15, 435 26, 435 19, 435 21, 435810, C12Q 140, C12Q 100

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active

049904450

ABSTRACT:
A stable reagent and kinetic assay for .alpha.-amylase is provided. The reagent comprises a substrate for the .alpha.-amylase and various enzymes and cofactors necessary to produce NADH as the end-product of a series of four reactions. The substrate, enzymes, and cofactors are provided in sufficient excess that the .alpha.-amylase from the test sample is the rate-limiting factor. The concentration of .alpha.-amylase in a test sample is determined by measuring the rate of increase in absorbance caused by the production of NADH. The use of the novel cofactor fructose-1,6-diphosphate has been found to impart improved stability to the reagent.

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