Sialidase from different sources compared for electrophoretically separating serum alkaline phosphatase fractions from liver and bone.
We compared sialidase (neuraminidase; EC 3.2.1.18) from Vibrio cholerae, Clostridium perfringens, and Arthrobacter ureafaciens, seeking to improve the electrophoretic separation of the liver and bone isoenzymes of alkaline phosphatase (EC 3.1.3.1) on cellulose acetate membranes. Resolution is decisively determined by the type and activity of sialidase used in the preincubation of serum sample. Sialidase from Arthrobacter ureafaciens is not suited for this method. For optimal separation of the two isoenzymes we recommend the use of sialidase from Vibrio cholerae, determination of its activity with a standard procedure such as described here (mucin or sialyl lactose as substrates), and a final concentration of sialidase activity of 2.0 or 2.9 U/L (measured with mucin or sialyl lactose) in the incubation mixture.
Department of Experimental Organ Transplantation, University Hospital Charité, Humboldt University Berlin, G.D.R.
K
K
Jung
M
M
Pergande
S
S
Klotzek
D000469:Alkaline Phosphatase / Q000097:blood*; D001173:Arthrobacter / Q000201:enzymology; D001842:Bone and Bones / Q000201:enzymology*; D003013:Clostridium / Q000201:enzymology; D004586:Electrophoresis / Q000379:methods; D006801:Humans; D006868:Hydrolysis; D007527:Isoenzymes / Q000097:blood; D008099:Liver / Q000201:enzymology*; D009439:Neuraminidase; D013045:Species Specificity; D014734:Vibrio cholerae / Q000201:enzymology
D007527:Isoenzymes; D000469:Alkaline Phosphatase; D009439:Neuraminidase