o-Dianisidine dihydrochloride → Hexose
Reaction equation
[OH:1][C@@H:2]1[O:10][C@H:9]([CH2:11][OH:12])[C@@H:7]([OH:8])[C@H:5]([OH:6])[C@H:3]1[OH:4].COC1C=C(C2C=CC(N)=C(OC)C=2)C=CC=1N.Cl.Cl>P([O-])([O-])([O-])=O.[Na+].[Na+].[Na+]>[CH2:11]([OH:12])[CH:9]1[O:10][CH:2]([OH:1])[CH:3]([OH:4])[CH:5]([OH:6])[CH:7]1[OH:8]Participants
4
Reagents, catalysts & solvents 1
Procedure
The assay was based on the method described by Sullivan and Ikawa (Biochimica et Biophysica Acta, 1973, 309:11-22), but modified to run in microtiter plates. An assay mixture contained 150 μl β-D-glucose (0.1 M in 0.1 M sodium phosphate buffer, pH 6.3), 120 μl 0.1 M sodium phosphate buffer, pH 6.3, 10 μl o-dianisidine-dihydrochloride (Sigma D-3252, 3.0 mg/ml in H2O), 10 μl peroxidase (POD) (Sigma P-8125, 0.1 ml in 0.1 M sodium phosphate buffer, pH 6.3) and 10 μl enzyme (HOX) solution. Blanks were made by adding buffer in place of enzyme solution.