GST融合蛋白纯化方法.docVIP

GST融合蛋白纯化方法?Purification?of?GST?Fused?Proteins Abstract: Many people have vented out frustration over insoluble GST-fused proteins. This is a protocol for enzymatically active soluble GST-fused proteins. All GST-fused proteins are rendered soluble with this technique though enzyme activitiy can range from 30-90%. Materials and Reagents STE Buffer 10 mM Tris-HCl, pH 8.01 mM EDTA150 mM NaCl Lysozyme solution 10 mg/ml in water (make fresh) PBS Elution Buffer 50 mM Tris.Cl, pH 9.020 mM GSH 10% Sarkosyl in STE Buffer 10% Triton X-100 in STE Buffer 1 M DTT 100 mM IPTG Procedure? ?Day