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文献和实验Thaw out 1 ml aliquot of fusion protein lysate. Centrifuge to remove precipitate. Incubate 1 ml lysate with 0.4 ml 50% GST beads. Incubate at 4C with mixing for 1 hour. Centrifuge 5K 2' in 1.5 ml eppendorf tubes. Wash the beads 4X with PBST
GST Fusion Protein Preparation
CLEAVAGEAlternate Mini PrepThaw out 1 ml aliquot of fusion protein lysate.Centrifuge to remove precipitate.Incubate 1 ml lysate with 0.4 ml 50% GST beads.Incubate at 4℃ with mixing for 1 hour.Centrifuge 5K 2' in 1.5 ml eppendorf tubes.Wash the beads 4X with PBST
Thrombin Cleavage of GST-Fusion protein
to cleave 1 mg of fusion protein at room temperature overnight. 1 X PBS buffer: 140mM NaCl; 2.7mM KCL; 10mM Na2HPO4; 1.8mM KH2PO4; pH 7.3 Thrombin Cleavage of Free Eluted Fusion protein First you have to calibrate amount of thrombin, temperature
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