Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to cleave > 95% of the terminal, β-D-galactose from 1 nmol Galβ1-4GlcNAcβ1- 3Galβ1-4Glc-7-amino-4-methyl-coumarin (AMC), in 1 hour at 37°C in a total reaction volume of 10 μl. Reaction Conditions
1X GlycoBuffer 1
Incubate at 37°C
1X GlycoBuffer 1
5 mM CaCl2
50 mM sodium acetate
(pH 5.5 @ 25°C)
Storage Buffer
50 mM NaCl
20 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
Heat Inactivation
65°C for 10 minutes Molecular Weight
Apparent: 231 kDa
Unit Assay Conditions
Two fold dilutions of β1-4 Galactosidase S are incubated with 1 nmol AMClabeled substrate and 1X GlycoBuffer 1 in a 10 μl reaction. The reaction mix is incubated at 37°C for 1 hour. Separation of reaction products are visualized via thin layer chromatography (1).