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Biotechniques ; 48(1): 61-4, 2010 Jan.
Article in English | MEDLINE | ID: mdl-20078429

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.


Subject(s)
Glutathione Transferase/isolation & purification , Inclusion Bodies/chemistry , Periplasmic Binding Proteins/isolation & purification , Recombinant Fusion Proteins/isolation & purification , Cholic Acids , Escherichia coli/genetics , Glutathione Transferase/analysis , Glutathione Transferase/genetics , Maltose-Binding Proteins , Octoxynol , Periplasmic Binding Proteins/analysis , Periplasmic Binding Proteins/genetics , Protein Folding , Recombinant Fusion Proteins/analysis , Recombinant Fusion Proteins/genetics , Sarcosine/analogs & derivatives , Solubility
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