Materials and Reagents

1. Antigen protein
   
2. PVDF or Nitrocellulose membrane (Amersham biosciences/GE Healthcare Dharmacon)
   
3. Ponceau S (Sigma-Aldrich, catalog number: P7170 )
   
4. non-fat Milk powder (Carnation, any your favorite brand in local store)
   
5. Tween 20 (Sigma-Aldrich)
   
6. Glycine
   
7. Acetic acid
   
8. 2 M Tris-HCl (pH 8.5)
   
9. NaCl
   
10. KCl
    
11. Na₂HPO₄
    
12. KH₂PO₄
    
13. Ultra pure water
    
14. TBST (see Recipes)
    
15. 10x TBS (1 L) (see Recipes)
    
16. PBS (pH 7.4) (see Recipes)
    
17. Ponceau S solution (see Recipes)
    

Equipment

1. Dialysis bag (Thermo Fisher Scientific/Pierce Antibodies, catalog number: 68035 )
   
2. Comb
   

Procedure