Materials and Reagents

1. Bacteria (e.g. Escherichia coli)
   
2. Sterile toothpicks or pipette tips
   
3. Circular Protran BA85 nitrocellulose membrane, 0.45 μm, 82 mm in diameter (Whatman, catalog number: 10401116 )
   
4. Albumin from bovine serum (BSA) (Sigma-Aldrich, catalog number: A7906 )
   
5. Tween 20 (Sigma-Aldrich, catalog number: P1379 )
   
6. Unconjugated primary antibody/antiserum (e.g. Polyclonal rabbit antiserum against Mat fimbriae of E. coli (Pouttu et al., 2001))
   
7. Alkaline phosphatase-conjugated secondary antibody (e.g. Alkaline phosphatase-conjugated swine anti-rabbit immunoglobulins (Dako, catalog number: D0306 ))
   
8. 1-Step BCIP/NBT solution (Pierce Antibodies, catalog number: 34042 )
   
9. Distilled water
   
10. Bacto Tryptone (BD Biosciences, catalog number: 211705 )
    
11. Bacto Yeast extract (BD Biosciences, catalog number: 212750 )
    
12. Bacto Agar (BD Biosciences, catalog number: 214010 )
    
13. 10% (w/v) SDS solution in distilled water
    
14. 0.5 M NaOH, 1.5 M NaCl in distilled water
    
15. 0.5 M Tris-HCl pH 7.5, 1.5 M NaCl in distilled water
    
16. 2x SSC (see Recipes)
    
17. LB agar plates(see Recipes)
    
18. Phosphate buffered saline (PBS) (see Recipes)
    
19. Blocking buffer: 2% (w/v) BSA in PBS (see Recipes)
    
20. Antibody dilution buffer: 1% (w/v) BSA in PBS (or the buffer as recommended in the antibody datasheet)(see Recipes)
    
21. Washing buffer: 0.05% (v/v) Tween 20 in PBS (see Recipes)
    

Equipment

1. 3 mm paper (Whatman 3MM, catalog number: 3030-6185 )
   
2. Petri dishes, 90 mm (Sterilin^®, catalog number: 101RT )
   
3. Beaker, 1,000 ml (Pyrex Corning, catalog number: 1000-1L )
   
4. Incubator (Termaks)
   
5. Belly dancer laboratory shaker (Stovall Life Science)
   
6. Sterile 18-gauge needle (BD Biosciences)
   
7. Tweezers (Bochem)
   

Procedure