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Detection of dextransucrase and levansucrase on polyacrylamide gels by the periodic acid-Schiff stain: Staining artifacts and their prevention

1986; Elsevier BV; Volume: 156; Issue: 2 Linguagem: Inglês

10.1016/0003-2697(86)90266-6

1096-0309

Arthur W. Miller, John F. Robyt,

Glycosylation and Glycoproteins Research

One use of the periodic acid-Schiff (PAS) stain is to detect dextransucrase and levansucrase activities on polyacrylamide gels by staining their polysaccharide products, dextran and levan. When gels with heavy dextran or levan bands were PAS stained, proteins other than dextransucrase and levansucrase also were stained, and a high background developed during storage. The staining of proteins other than dextransucrase and levansucrase is caused by the diffusion of the periodateoxidized carbohydrate before and after staining. This diffusion could be greatly slowed, and the staining artifact decreased, by following the PAS stain by a crosslinking treatment of the carbohydrate-dye complex. Protein staining artifacts could be prevented by using chymotrypsin to remove the protein from the gel at the stage after polysaccharide synthesis but before the PAS stain.