Description
Alpha Galactosidase from E. coli cleaves α(1-3)- and α(1-6)-linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins. There is no activity on α(1-4) linked galactose. It is particularly efficient for removing α-linked galactose under conditions where the pH must be neutral or above, for example, with living cells.Molecular Weight ~80,000 daltonsContents Alpha galactosidase in 50 mM sodium phosphate, pH 7.5 included with 20 µL and 60 µl pack sizes: Reaction buffer - 250mM Sodium phosphate, pH 6.5Specificity Non-reducing terminal alpha-(1-3)- and alpha-(1-6)- galactose. There is no activity on alpha-(1-4)-galactose.Stability Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.Specific Activity One unit of alpha-(1-3,6) Galactosidase is defined as the amount of enzyme required to produce 1 µmole of p-nitrophenol (pNP) in 1 minute at 25°C pH 6.5 from p-nitrophenyl-alpha-D-galactopyranoside. Purity α(1-3,6) galactosidase is tested for contaminating protease as follows; 10 µg of denatured BSA is incubated for 24 hours at 37°C with 2 µL of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation. The production host strain has been extensively tested and does not produce any detectable glycosidases.of the BSA band after SDS-PAGE should show no evidence of degradation. Directions for use 1. Add up to 100 µg of asialoglycoprotein or 1 nmol of oligosaccharide to tube. 2. Add water to 13 µl and 4 µl 5X Reaction Buffer. 3. Add 2 µl alpha-(1-3,6)-Galactosidase. 4. Incubate at 37°C for 1 hour. Longer incubations are necessary if fucose is present on the penultimate sugar.Applications Structural analysis of oligosaccharides Xenograft transplantation studies Removing heterogeneity from glycoproteins