Identification of glycoproteins on nitrocellulose membranes and gels.
In this article we describe a procedure for the detection of glycoproteins on gels employing the periodic acid-Schiff's reagent. In addition, a number of staining protocols and direct binding ELISA, employing antibodies and lectins, are described for the identification and quantitation of glycoproteins after their immobilization by dot, slot, or Western blotting onto nitrocellulose membranes. We document, in detail, the conditions (i.e., the effect of solvent and detergents) for the immobilization of one specific family of O-linked glycoproteins, namely mucins. However, taking into account our suggestions, these procedures should be applicable to other types of glycoprotein.
Thornton
D J
DJ
Wellcome Trust Centre for Cell-Matrix Research, University of Manchester, School of Biological Sciences, UK.
Carlstedt
I
I
Sheehan
J K
JK
D000180:Acrylic Resins; D003101:Collodion; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005782:Gels; D006023:Glycoproteins / Q000032:analysis*; D010503:Periodic Acid-Schiff Reaction; D012685:Sepharose; D013194:Staining and Labeling
D000180:Acrylic Resins; D005782:Gels; D006023:Glycoproteins; C016680:polyacrylamide gels; D003101:Collodion; D012685:Sepharose