Expression and Purification of GST-tagged Proteins from E. coli

Materials and Reagents

1. Escherichia coli (E. coli) strain of choice
   
2. IPTG (Sigma-Aldrich)
   
3. Imidazaole (Sigma-Aldrich)
   
4. Protease inhibitor (Roche Diagnostics)
   
5. Lysozyme (Sigma-Aldrich)
   
6. Glutothion sepharose 4B beads (Thermo Fisher Scientific/Pierce Antibodies, catalog number: 15160 )
   
7. Glutathione (Thermo Fisher Scientific/Pierce Antibodies, catalog number: 78259 )
   
8. NP-40 (Sigma-Aldrich)
   
9. Bacto trypton
   
10. Yeast extract
    
11. Liquid nitrogen
    
12. NaCl
    
13. KCl
    
14. Na₂HPO₄
    
15. KH₂PO₄
    
16. Protease inhibitor
    
17. Lysis buffer (see Recipes)
    
18. 1x PBS buffer (see Recipes)
    
19. Elution buffer (see Recipes)
    
20. 2x YT medium (see Recipes)
    

Equipment

1. Sonicator
   
2. Centrifuges
   
3. Pellete
   
4. Water bath
   

Procedure