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Purifying natively folded proteins from inclusion bodies using sarkosyl, Triton X-100, and CHAPS

Authors :
Hu Tao
Wenjun Liu
Brandi N. Simmons
Helen K. Harris
Timothy C. Cox
Michael A. Massiah
Source :
BioTechniques, Vol 48, Iss 1, Pp 61-64 (2010)
Publication Year :
2010
Publisher :
Future Science Ltd, 2010.

Abstract

We describe a rapid, simple, and efficient method for recovering glutathione S-transferase (GST)- and His6-tagged maltose binding protein (MBP) fusion proteins from inclusion bodies. Incubation of inclusion bodies with 10% sarkosyl effectively solubilized >95% of proteins, while high-yield recovery of sarkosyl-solubilized fusion proteins was obtained with a specific ratio of Triton X-100 and CHAPS. We demonstrate for the first time that this combination of three detergents significantly improves binding efficiency of GST and GST fusion proteins to gluthathione (GSH) Sepharose.

Details

Language :
English
ISSN :
19409818 and 07366205
Volume :
48
Issue :
1
Database :
Directory of Open Access Journals
Journal :
BioTechniques
Publication Type :
Academic Journal
Accession number :
edsdoj.195fae27bbf8457fbd0a551939110b8b
Document Type :
article
Full Text :
https://doi.org/10.2144/000113304