Using neuraminidase (Product No. 10269611001) to remove sialic acid from purified lipooligosaccharide (LOS)
The enzyme can be used to cleave sialic acids from proteins. A mixture of N-glycosidase F, O-glycosidase, and neuraminidase is often useful (O-glycans often have sialidized structures that could be hydrolyzed by neuraminidase and then O-glycosidase could continue hydrolyzing the structure). The enzyme/substrate ratio should be approx. 0.04 U/25 - 80 μg.
To use neuraminidase from Vibrio cholerae (instead of neuraminidase from Arthrobacter) together with O-glycosidase under the same conditions, consider the pH-optima of O-glycosidase and the V. cholerea neuraminidase, and therefore, use pH 6 instead of pH 7.2.
The following procedure adapted from Corral, et al., (1990) BBRC 172:1349-1356:
Specific inhibitors of neuramindase (silaidase) from Vibrio cholerae:
It has not been determined whether SDS, guanidine HCl, or urea inhibit the enzyme, but inactivation is possible.
It has not been determined whether CHAPS, Triton X100/X114, 2-mercaptoethanol, or Manidet P40 inhibit the enzyme, but most likely no inhibition occurs. No information is available on inhibition by DTT.
如要继续阅读,请登录或创建帐户。
暂无帐户?