Abstract
An assay method for elastase activity was established using a natural substrate, elastin, because synthetic low molecular weight substrates may not reflect elastase activity accurately. A fluorometric assay method with fluorescamine was used, after a deproteinizing process with trichloroacetic acid. Neutralization of 5% trichloroacetic acid was done with 0.2N NaOH, and the fluorescence of the products formed from elastin by elastase with fluorescamine was determined in dioxane-phosphate buffer after the enzymatic reaction had been carried out in H3BO4-NaOH buffer, pH 8.8, with 10 mg elastin/1.1 ml of assay mixture. As little as 1.56×10-5 mg of elastase can be determined by the present method, and there was a good correlation between the results of a conventional method using succinyl alanylalanylalanyl-p-nitroanilide and the present method.
