Neuraminidase Cp cleaves all non-reducing terminal non-branched α(2-3) and α(2-6) sialic acid residues from complex carbohydrates and glycoproteins.
recombinant from C. perfringens
Part Number – Amount of Enzyme
E-S005 – 60 µLs¹
E-S005-20 – 20 µLs¹
E-S005-200 – 200 µls²
¹ includes buffer
² includes enzyme only
$198 – $1,377
Sialidase, N-acetylneuraminate glycohydrolase, Exo-alpha-sialidase
α(2-3,6) Neuraminidase Cp cleaves all non- reducing terminal non-branched &alpha(2-3)- and α(2-6) sialic acid residues from complex carbohydrates and glycoproteins. There is no detectable activity on α (2-8) or α(2-9) linkages or on branched α(2-3) or α(2-6) linkages. The relative cleavage rates for different linkages are: α(2-3) > α(2-6).
α(2-3,6) Neuraminidase Cp will not cleave branched sialic acids (linked to an internal residue). Use α(2-3,6,8,9) Neuraminidase (E-S001) for α(2-8) or branched sialic acids. To cleave only non-reducing terminal α(2-3) unbranched sialic acid residues, use α(2-3) Neuraminidase (E-S007).
α(2-3,6) Neuraminidase Cp is isolated from a clone of Clostridium perfringens. The enzyme has been extensively characterized using oligosaccharide standards.
Source recombinant from Clostridium perfringens in E. Coli.
Neuraminidase Cp in 20 mM Tris-HCl, 25 mM NaCl, pH 7.5
Included its 20 µL and 60 µL pack sizes:
5x Reaction Buffer 250 mM sodium phosphate, pH 6.0
Specific Activity >250 U/mg
Activity 15 U/ml
Molecular weight 41,000 daltons
pH range 50 mM sodium phosphate (pH 6.0) provides the optimal buffer for enzyme activity with 3’-siayllactose, a standard substrate. If glycosidase treatment is performed at suboptimal pH because of glycoprotein solubility or activity requirements, expect some diminution in enzyme activity.
1. Add up to 100 μg of glycoprotein or 1 nmol of oligosaccharide to tube.
2. Add water to 14 μL
3. Add 4 μL 5X Reaction Buffer.
4. Add 2 μL α (2-3,6) Neuraminidase.
5. Incubate at 37°C for 1 hour.
Desialylation may be monitored by SDS-PAGE if the size differential between native and desialylated protein is sufficient for detection.
Specifictity Cleaves all nonreducing terminal non-branched alpha-(2-3) and alpha-(2-6)-sialic acid residues from complex carbohydrates and glycoproteins.
Relative cleavage rates for different linkages are: (2-3) > (2-6).
Specific Activity Assay 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-methylumbelliferyl)-alpha-D-N-acetylneuraminic acid].
Storage Store enzyme at 4˚C.
Neuraminidase CP References
Corfield, A. P., H. Higa, J. C. Paulson and R. Schauer. The specificity of viral and bacterial sialidases for alpha(2-3) and alpha(2-6)-linked sialic acids in glycoproteins. Biochem Biophys Acta 744: 121-12 6 (1983).
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 (1993).
Kobata, A. Use of endo- and exoglycosidases for structural studies of glycoconjugates. Anal Biochem 100: 1-14 (1979).
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).
Uchida, Y., Y. Tsukada and T. Sugimori. Enzymatic properties of neuraminidases from Arthrobacter ureafaciens. J Biochem (Tokyo) 86: 573-58 5 (1979).
Roggentin, P, B. Rothe, F Lottspeich and R. Schauer. Cloning and sequencing of a Clostridium perfringens sialidase gene. FEBS Lett 238: 31-34 (Se pt 1988).
Roggentin P., R.G . Kleineidam and R. Schauer. Diversity in the properties of two sialidase isoenzymes produced by Clostridium perfringens spp. Biol Chem Hoppe-Seyler 376: 569-575 (1995)