Staining of glycoproteins in polyacrylamide and agarose gels with fluorescent lectins

Furlan, M.; Perret, B.A.; Beck, E.A.

Analytical Biochemistry 96(1): 208-214


ISSN/ISBN: 0003-2697
PMID: 91330
DOI: 10.1016/0003-2697(79)90574-8
Accession: 006471754

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A simple and sensitive method for staining of the carbohydrate moiety of glycoproteins in polyacrylamide or agarose electrophoretic gels is described. Gels are incubated in a solution of fluorescein-labeled concanavalin A. Following destaining with a neutral buffer, glycoproteins exhibit fluorescence under long-range UV light. The glucose/mannose containing .beta.- and .gamma.-chains of human fibrinogen give fluorescent bands but the carbohydrate-free .alpha.-chain does not react. Less than 100 ng of hexose bound to fibrinogen .beta.- or .gamma.-chains could be detected. The procedure is suitable for staining of other carbohydrate residues in glycoproteins which can be recognized by specific agglutinins, as shown by binding of fluorescein-labeled lectins from Ricinus communis to galactose residues of fibrinogen.