Promega Corporation

Detection of Protein:Protein Interactions Using the MagneGST™ Pull-Down System

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Abstract

The MagneGST™ Pull-Down System was designed to detect protein interactions between GST-fusion proteins expressed in E. coli lysates and prey proteins expressed in vitro in the TnT® T7 Quick Coupled Transcription/Translation System. The MagneGST™ System provides glutathione (GSH)-linked magnetic particles that allow immobilization of GST-fusion proteins (bait) from bacterial lysates. The magnetic nature of the particles allows easy and quick processing of multiple samples and provides enhanced recovery. Prey protein synthesized in the TnT® T7 Quick Coupled Transcription/Translation reaction is captured using bait protein (GST-fusion protein) immobilized on MagneGST™ (GSH) Particles. Nonspecifically bound proteins are washed away, and the prey protein is analyzed. We tested the method described with two different protein:protein interaction systems and detected the results by either radioactive labeling or Western blotting.

Promega Notes 87, 23–26.

Marjeta Urh, Jacqui Sankbeil, Don Creswell and Dan Simpson

Promega Corporation
2004

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