糖蛋白染色比较.pdfVIP

ANALYTICAL BIOCHEMISTRY 73, 192- 197 (1976) A Sensitive Fluorescent Method for the Detection of Glycoproteins in Polyacrylamide Gels1 ALLEN E. ECKHARDT, COLLEEN E. HAYES, AND IRWIN J. GOLDSTEIN Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109 Received November 21, 1975; accepted January 22, 1976 A sensitive method is described for the detection of glycoproteins after polyacrylamide gel electrophoresis. Dansyl hydrazine is condensed with aldehydic functions generated by periodate oxidation of protein bound carbohydrate. The resulting hydrazones are reduced to the stable hydrazine derivatives with sodium borohydride. After destaining, the fluorescent-labeled glycoproteins are visualized with a long-range uv light source. The dansyl hydrazine staining tech- nique was shown to detect less than 40 ng of carbohydrate bound to Bandeiraea simplicifolia lectin. The principle methods for the detection of glycoproteins in polyacryl- amide gels are the periodic acid-Schiff stain (PAS) (1,2,4) and the Alcian blue stain (3). Both of these procedures involve the fixation of the glyco- proteins in the gel followed by oxidation of sugar residues with periodic acid generating aldehydic functions, reduction of the excess periodate with sodium metabisulfite and staining with either basic fuchsin or Alcian blue. The minimum amount of pr