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

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

  • Biotechniques. 2010 Jan;48(1):61-4. doi: 10.2144/000113304.
Hu Tao 1,  Wenjun Liu,  Brandi N Simmons,  Helen K Harris,  Timothy C Cox,  Michael A Massiah
Affiliations

Affiliation

  • 1 Department of Biochemistry and Molecular Biology, Oklahoma State University, Stillwater, OK 74078, USA.
PMID: 20078429 DOI: 10.2144/000113304
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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