Literature DB >> 20078429

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

Hu Tao1, Wenjun Liu, Brandi N Simmons, Helen K Harris, Timothy C Cox, Michael A Massiah.   

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.

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Year:  2010        PMID: 20078429     DOI: 10.2144/000113304

Source DB:  PubMed          Journal:  Biotechniques        ISSN: 0736-6205            Impact factor:   1.993


  33 in total

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