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Methods Mol Biol. 2019;1893:273-280. doi: 10.1007/978-1-4939-8910-2_20.

GST Pull-Down Assay to Measure Complex Formations.

Author information

1
Structural Biology Laboratory, Nara Institute of Science and Technology, Nara, Japan.
2
Structural Biology Laboratory, Nara Institute of Science and Technology, Nara, Japan. hakosima@bs.naist.jp.

Abstract

The GST pull-down assay is an intuitive and fast in vitro method for analyzing protein-protein or protein-ligand interactions and is comprised of a "bait" which is a GST-fused protein expressed in E. coli host or a baculovirus expression system and a "prey" which comprises putative binding partner protein(s) or other ligand molecule(s). This method is suitable for examining the direct interaction between two purified proteins and estimating the extent of the affinity.

KEYWORDS:

Direct protein–protein interaction; GST pull-down assay; Glutathione S-transferase; Glutathione-conjugated resin; In vitro assay; Screening of binding partners

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