Alpha Galactosidase from E. coli cleaves α(1-3)- and α(1-6)-linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins.
recombinant from E. coli in E. coli
Part Number – Amount of Enzyme
E-AG02 – 60 µLs¹
E-AG02-20 – 20 µLs¹
E-AG02-200 – 200 µls²
¹ includes buffer
² includes enzyme only
α-D-galactoside galactohydrolase, melibiase.
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.
Source recombinant from E. coli in E. coli
Alpha Galactosidase enzyme in 50 mM sodium phosphate, pH 7.5
Included with 20 µL and 60 µL pack sizes:
5x Reaction Buffer – 250 mM sodium phosphate, pH 6.5
Specific Activity >30 U/mg
Activity 80 U/ml
Molecular weight ~80,000 daltons
1. Add up to 100 μg of glycoprotein or 1 nmol of oligosaccharide to tube.
2. Add water to 13 μL
3. Add 4 μL 5X Reaction Buffer.
4. Add 2 μL α(1-3,6) galactosidase.
5. Incubate at 37°C for 1 hour.
NOTE: Longer incubations are necessary if fucose is present on the penultimate sugar.
Specifictity Cleaves Alpha-(1-3)-and Alpha-(1-6)-linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins.
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.
Storage Store enzyme at 4˚C.
Purity Each lot of Alpha Galactosidase is tested for contaminating protease as follows; 10 μg of denatured BSA is incubated for 24 hours 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.
Stability Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.
Alpha Galactosidase References
1. Kobata, A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem 100: 1- 14 (1979).
2. Prime, S. J. Dearnley, A.M. Venton, R.B. Parekh and C.J. Edge. Oligosaccharide sequencing based on exo- and endoglycosidase digestion and liquid chromatographic analysis of the products. J Chromatogr A 720: 263-274 (1996)
3. Dwek, R.A. , C.J. Edge, D.J. Harvey, M.R. Wormald and R.B. Parekh. Analysis of glycoprotein-associated oligosaccharides. Ann Rev Biochem 62: 65-100.