NEURAMINIDASE (SIALIDASE) F. V.CHOLERAE

General description

Neuraminidase is an acylneuraminyl hydrolase which hydrolyzes terminal N- or O-acylneuraminic acids which are α2,3-, α2,6-, or α2,8-linked (rate: α2,6 > α2,3 > α2,8) to oligosaccharides, polysaccharides, mucopolysaccharides, glycoproteins, and glycolipids. Noteworthy, for the hydrolysis of glycolipids, the presence of a detergent is necessary. Because of the broad substrate specificity, the enzyme is very well suited for the complete removal of sialic acids from glycoconjugates of a wide variety of biological materials (e.g., in cytology, on cell surfaces, viruses etc.).

Specificity

Hydrolyzes terminal N- or O-acyl-neuraminic acids that are α2,3-, α2,6-, or α2,8-linked to galactose, Hex, NAc, or N- or O-acylated neuraminyl residues in oligosaccharides/glycoconjugates or colominic acid. Relative rate of cleavage is α2,3 >α2,6 >α2,8, determined on bonds in tri- and tetrasaccharides.

Application

Neuraminidase has been used:

• to remove cis-acting sialic acids in CHO (chinese hamster ovary) cells
• for deglycosylation studies

Unit Definition

One unit is the enzyme activity that hydrolyzes 1 μmol N-acetyl-neuraminosyl-D-lactose within 1 min at +37 °C under the following incubation conditions:
10 mM N-acetyl-neuraminosyl-D-lactose, 50 mM sodium acetate, 4 mM calcium chloride, bovine serum albumin, 100 μg/ml, pH 5.5. The activity is determined by measuring the released D-lactose using the β-galactosidase/galactose dehydrogenase method. Under the same conditions, 1 μmol N-acetylneuraminic acid per min is split off from human acid α₁-glycoprotein (10 mg/ml incubation mixture) by 1 U neuraminidase. Released N-acetyl-neuraminic acid can be determined using, for example, the thiobarbituric acid method.

Physical form

Solution in 50 mM sodium acetate, 154 mM sodium chloride, 9 mM calcium chloride, 0.1% Micr-O-Protect (w/v), human serum albumin, 25 mg/l, pH 5.5. The preparation contains 10 mM EDTA.
Note: The serum used for this preparation was tested for HBs antigen and for the presence of antibodies to HIV-1, HIV-2, HCV, and found to be negative, according to the current quality control procedures.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Legal Information

The sale of the Product does not exhaust or grant any rights in third party patents including patents of companies of the F. Hoffmann - La Roche AG group of companies, in particular, for the use of modified antibodies obtained by using the product.