Description
α(2-3) Neuraminidase Sp cleaves exclusively the non-reducing terminal α(2-3) unbranched sialic acid residues from complex carbohydrates and glycoproteins. There is no detectable activity on α(2-6) or α(2-8) linkages or on branched α(2-3) linkages. To cleave all non-reducing terminal sialic acid residues including branched sialic acids (linked to an internal residue) from complex carbohydrates and glycoproteins, use α(2-3,6,8,9) Neuraminidase Au.ContentsNeuraminidase Sp in 50 mM sodium phosphate, pH 7.5Included with 20 µL and 60 µL pack sizes:5x Reaction Buffer 250 mM sodium phosphate, pH 6.0SpecificityAll non-reducing terminal branched and unbranched a-(2- 3) sialic acid.Specific Activity AssayOne unit of Neuraminidase is defined as the amount of enzyme required to produce 1 µmole of methylumbelliferone in 1 minute at 37°C, pH 5.0 from MU-NANA (2’.-(4-methyl-umbelliferyl)-alpha-D-N acetylneuraminic acid].Molecular Weight~75,000 daltonspH optimum6.0, active over the range 4.5-7.50 mM sodium phosphate (pH 6.0) provides the optimal buffer for enzyme activity with sialyllactose, a standard substrate. If glycosidase treatment is performed at suboptimal pH because of glycoprotein solubility or activity requirements, expect some diminution in enzyme activity.FormulationThe enzyme is provided as a sterile-filtered solution in in 50 mM Sodium phosphate pH 7.5.StabilityStable at least 12 months when stored properly. Several days exposure to ambient temperatures will not reduce activity.PurityNeuraminidase Sp 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.The production host strain has been extensively tested and does not produce any detectable glycosidases.Directions for use1. Add up to 100 µg of glycoprotein or 1 nmol of oligosaccharide to tube.2. Add de-ionized water to a total of 14 µl.3. Add 4 µl 5x Reaction Buffer 6.0.4. Add 2 µl Neuraminidase Sp.5. Incubate at 37°C for 1 hour.NOTE: longer incubation times are necessary if branched sialic acids are present.Desialylation may be monitored by SDS-PAGE if the size differential between native and de-sialylated protein is sufficient for detection. Applications•Structural analysis of oligosaccharides•Determining sialic acid linkage•Glycoprotein deglycosylation•Removing heterogeneity from glycoproteins