GST tag removal Protocol

GST tag Removal using GE Healthcare's PreScission Protease

GST tags may be removed using GE Healthcare’s PreScission Protease. The PreScission Protease is a genetically engineered fusion protein consisting of human rhinovirus 3C protease and GST. This protease was specifically designed to facilitate removal of the protease by allowing simultaneous protease immobilization and cleavage of GST-tagged proteins produced from the pGEX-6P vectors (pGEX-6P-1, pGEX-6P-2, and pGEX-6P-3). PreScission Protease specifically cleaves between the Gln and Gly residues of the recognition sequence of LeuGluValLeuPheGlnGlyPro. The protease has maximal activity at 4C, and cleavage can thus be performed at low temperatures, improving the stability of the target protein. One unit will cleave > 90% of 100ug of a test GST-tagged protein in 50 mM Tris-HCl, 150 mM sodium chloride, 1 mM EDTA, 1 mM DTT, pH 7.0 at 5C for 16 h.