Beta-(1-3,4,6) Galactosidase

Product Description Beta-(1-3,4,6) Galactosidase cleaves all β(1-3) and β(1-4) linked non-reducing, terminal galactose. β(1-6) linked galactose is released at a slower rate. The enzyme is a glycoprotein. β(1-4) galactose is by far the most common linkage found in N-linked oligosaccharides. For cleavage of this linkage we recommend β(1-4) Galactasidase. Contents 200 µls of Beta-(1-3,4,6) Galactosidase in 20 mM Tris-HCl, 50 mM NaCl, 0.5mg/ml BSA,pH 7.5 5x Reaction Buffer- 500 mM sodium citrate/phosphate pH 4 Optimum pH 4 The supplied buffer concentrate provides the optimal pH for enzyme activity with the standard substrate. If glycosidase treatment is performed at suboptimal pH because of glycoprotein solubility or activity requirements, expect some diminution in enzyme activity. Specific Activity One unit of ß-(1-3,4,6)-Galactosidase is defined as the amount of enzyme required to produce 1 µmole of p-nitrophenol (pNP) in 1 minute at 37°C, pH 4.0 from p-nitrophenyl-ß-D-galactopyranoside. Specificity Cleaves all ß1-3 and ß1-4 linked non-reducing, terminal galactose. ß1-6 linked galactose is released at a slower rate. Formulation The enzyme is provided as a sterile-filtered solution in 20 mM Tris-HCl pH 7.5, 50 mM NaCl, 0.5 mg/ml BSA, pH 7.5. Stability Stable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity. Purity ß-(1-3,4,6)-Galactosidase is tested for contaminating protease as follows: 10 µg of denatured BSA is incubated at 37°C for 24 hours with 2 µl of enzyme. SDS-PAGE analysis of the treated BSA shows no evidence of degradation. Each lot is also tested for contaminating activities by incubating the enzymes with the appropriate substrates for 24 hours; the detection limit is 5 µU/ml (IUB). A passing lot will have no detectable activity. Directions for use 1. Add up to 100 µg of asialoglycoprotein or 1 nmol of oligosaccharide to tube. 2. Add deionized water to a total of 14 µl. 3. Add 4 µl of 5x Reaction Buffer 4. 4. Add 2 µl ß-Galactosidase. 5. Incubate at 37°C for 1 hour. For glycoproteins, cleavage may be monitored by SDSPAGE if the size differential between native and degalactosylated protein is sufficient for detection

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