Staining of Concanavalin A‐reactive glycoproteins on polyacrylamide gels with horseradish peroxidase — a critical evaluation

Abstract

Abstract The method to localize and quantitate glycoproteins on polyacrylamide gels is based on the binding of Concanavalin A to the glycoproteins and the subsequent coupling of horseradish peroxidase to the glycoprotein ‐ Concanavalin A complex. The complex is visualized by the oxidation of 3,3 ‐diaminobenzidine with horseradish perexidase ‐ H 2 O 2 , forming a brownish stain at the gel surface. Various parameters are analyzed and a scheme for the optimal staining of macro and micro slab gels is given. The quantitative evaluation of the stained gels is demonstrated. The staining and quantitation of nitrocellulose transfers is also shown.