Silver staining of extensively glycosylated proteins on sodium dodecyl sulfate-polyacrylamide gels: enhancement by carbohydrate-binding dyes.
Two methods are described for detecting less than 1 microgram of highly glycosylated proteins, such as mucins, on sodium dodecyl sulfate-polyacrylamide gels. They combine commonly employed periodic acid-Schiff (PAS) and Alcian blue dyes with silver stain. Carbohydrate prestaining renders mucins more cationic and favors greater complexation with ionic silver. Comparisons of different mucin samples stained either with PAS-silver or alcian blue-silver indicate differential staining between the two techniques. Such differences may, in part, be due to an affinity of Alcian blue for sulfated glycoproteins. These two staining protocols when used in conjunction with silver staining alone are particularly valuable for assessing sample purity and for detecting contaminating proteins during mucin purification protocols.
Jay
G D
GD
Department of Pathology, State University of New York, Stony Brook 11794.
Culp
D J
DJ
Jahnke
M R
MR
D000423:Alcian Blue / Q000378:metabolism; D000818:Animals; D050260:Carbohydrate Metabolism; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D009077:Mucins / Q000378:metabolism; D010503:Periodic Acid-Schiff Reaction; D011509:Proteoglycans / Q000378:metabolism; D051381:Rats; D012834:Silver / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate; D013194:Staining and Labeling; D013552:Swine; D014168:Transferrin / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000378:metabolism
D006023:Glycoproteins; D009077:Mucins; D011509:Proteoglycans; D014168:Transferrin; D000509:alpha-Fetoproteins; D012967:Sodium Dodecyl Sulfate; D012834:Silver; D000423:Alcian Blue