Product Description 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. For cleaving Beta Galactosidase we recommend ß-(1-4) Galactosidase, recombinant from Streptococcus pneumoniae or ß-(1,3,4,6) Galactosidase, purified from bovine testes. Specifictity Cleaves Alpha-(1-3)-and Alpha-(1-6)-linked, non-reducing terminal galactose from complex carbohydrates and glycoproteins.
Source recombinant from E. coli in E. coli
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.
EC 3.2.1.22 Specific Activity >30 U/mg Activity 80 U/ml Molecular weight ~80,000 daltons
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.
Contents 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
Suggested Usage 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.
Storage Store enzyme at 4˚C. Stability Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.
The production host strain has been extensively tested and does not produce any detectable glycosidases.
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