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Neuraminidase (Sialidase) Protocol

Product No. 11080725001

Protocol

Neuraminidase (Sialidase) from Vibrio cholerae: Removal of sialic acid from purified lipooligosaccharide (LOS)

The optimal pH for the enzyme is 5.5 to 6.2.
In some cases, it helps to denature the protein to make sites more accessible.
Below are suggested incubation conditions for the deglycosylation of glycoproteins, and denaturation conditions.

The enzyme can be used to cleave sialic acids off proteins:

A mixture of N-glycosidase F, O-glycosidase, and neuraminidase is often used; O-glycans typically have sialized structures which can be hydrolyzed by neuraminidase, and then O-glycosidase can be use to further hydrolyze O-glycans.

A useful enzyme/substrate ratio is approx. 0.04 U/25 - 80 μg.

Combination with O-glucosidase:

When using neuraminidase from Vibrio cholerae, instead of neuraminidase from Arthrobacter, together with O-glycosidase, take into account the pH-optima of O-glycosidase and Vibrio neuraminidase, and work at pH 6 instead of pH 7.2.

Materials

     
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