Selective Staining of Platelet Glycoproteins Using Two-Dimensional O’Farrell Gel Electrophoresis and Avidin-Biotin-Conjugated Lectins

Summary

A time-saving and sensitive high resolution method lor the analysis and identification of platelet glycoproteins using non-radioactive compounds has been developed. Platelet proteins (50 μg) of normal subjects were separated by isoelectric focusing and SDS-PAGE. Proteins were either stained with silver or electroblotted onto nitrocellulose. Nitrocellulose blots were treated with the following biotinylated lectins : Abrus precatorius lectin, concanavalin A, Lens culinaris lectin, or wheat germ agglutinin. Staining was achieved by avidin-biotin-coupled peroxidase using 4-chloro-l-naphthol as the substrate. A rapid overview of platelet glycoproteins may be obtained by applying all the lectins to a single blot.

Keywords

Platelet glycoproteins - Two-dimensional O’Farrell gel electrophoresis - Western blot - Avidin-biotin - Lectins

• References

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