Materials and Reagents

1. Bacterial strain (S. mutans UA159 wild-type strain or other S. mutans strains)
   
2. Todd-Hewitt broth (BD Biosciences)
   
3. Yeast-Extract (BioShop)
   
4. Tris Base
   
5. NaCl
   
6. Sodium dodecyl sulfate (SDS)
   
7. Triton X-100
   
8. KOH
   
9. MgCl₂
   
10. Glycine
    
11. Glycerol
    
12. Bromophenol blue
    
13. Methylene blue
    
14. 40% Acrylamide/Bis solution (37.5:1 acrylamide:bisacrylamide) (BioShop)
    
15. Ammonium persulfate (Sigma-Aldrich)
    
16. TEMED (BioBasic, Inc.)
    
17. Ethanol
    
18. Isopropanol
    
19. dH₂O
    
20. Filter paper
    
21. Precision Plus Protein Prestained Standards (Bio-Rad Laboratories)
    
22. THYE broth (see Recipes)
    
23. 20 mM Tris, 100 mM NaCl (pH 7.4) (see Recipes)
    
24. 1.5 M Tris (pH 8.8) (see Recipes)
    
25. 0.5 M Tris (pH 6.8) (see Recipes)
    
26. SDS-PAGE loading buffer (see Recipes)
    
27. Tris-Glycine SDS running buffer (see Recipes)
    
28. Zymogram renaturing buffer (see Recipes)
    
29. Staining solution (see Recipes)
    
30. 10% separating gel solution (see Recipes)
    
31. 4% stacking gel solution (see Recipes)
    

Equipment

1. 15-ml canonical tubes
   
2. Flasks
   
3. 1.5 ml microcentrifuge tubes
   
4. Refrigerated centrifuge
   
5. Refrigerated microcentrifuge
   
6. CO₂ incubator
   
7. Spectrophotometer
   
8. Disposable plastic cuvettes
   
9. Protein mini gel cassettes
   
10. Heating block module
    
11. Power supply
    
12. Orbital shaker
    
13. 37 °C temperature chamber
    

Procedure