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Detection of Protein:Protein Interactions Using the MagneGST™
Pull-Down System
Marjeta Urh, Ph.D., Jacqui Sankbeil, M.S., Don
Creswell, B.S., and Dan
Simpson, Ph.D.
Promega Corporation
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.
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