Enzyme determination method

Details Enzyme determination method Enzyme determination method

1976-11-19

1979-04-24

Naff, David M.

Textiles: fiber preparation

1951035C, G01N 3114

Patent

active

041510437

ABSTRACT:
An improved colorimetric aldolase method in which the quantity of aldolase in a sample is determined by the rate at which it enzymatically cleaves fructose-1, 6-diphosphate (FDP) into glyceraldehyde-3-phosphate (GAP) and dihydroxyacetone phosphate (DHAP); the phosphate group is hydrolyzed from the GAP and DHAP; and one of the hydrolysis products is measured. The initial enzymatic reaction is carried out in a reaction mixture containing a concentrated tris-(hydroxymethyl)-aminomethane buffer, pH 7.0, and a small quantity of sodium fluoride, but without added hydrazine or other compound to prevent the enzymatic conversion of a great majority of GAP to DHAP in the reaction mixture. Hydrolysis of the reaction products and conversion to a colored reaction product is carried out by a procedure which yields intense, long-lasting, reproducible color. An improved standard includes dihydroxyacetone carried through the same procedure.

REFERENCES:
sibley et al., J. Biol. Chem., 177, pp. 859-872 (1949).
Dounce et al., J. Biol. Chem., 185, pp. 769-780 (1950).
Riddle et al., J. Biol. Chem., 243, pp. 2718-2724 (1968).

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