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Updated: Sep 14, 2025

Identification of Protein Interacting Partners Using Tandem Affinity Purification
Published on: February 25, 2012
Protocol to isolate glycoproteins from cultured mammalian cells and tissue samples using wheat germ agglutinin
Simran Chopra1, Otsuware Linda-Josephine Kadiri1, Kai Kappert2
1Department of Molecular and Experimental Nutritional Medicine, Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558 Nuthetal, Germany.
Abstract:
Wheat germ agglutinin (WGA) precipitation enables selective enrichment of glycoproteins such as density-enhanced phosphatase (DEP-1) from protein lysates. Here, we present a protocol to isolate glycoproteins from cultured mammalian cells and tissue samples using WGA agarose beads. We describe steps for lysate preparation, bead incubation, and sequential washing to remove unbound proteins, followed by elution and denaturation. We then detail procedures for SDS-PAGE and western blot analysis to assess glycoprotein enrichment and binding specificity. For complete details on the use and execution of this protocol, please refer to Chopra et al.1.

