Materials and Reagents

1. Oligo DNA 5’ end-labeled with IRDye 700 or IRDye 800 (sense strand) (Integrated DNA Technologies)
   
2. Non-labelled oligo DNA (both sense and antisense strands)
   
3. Non-labelled mutated oligo DNA (both sense and antisense strands)
   
4. Recombinant DNA-binding proteins expressed in Escherichia coli (E. coli) (10 ng/µl in protein storage buffer)
   
5. Sterile distilled water (SDW)
   
6. Odyssey infrared EMSA kit (LI-COR, catalog number: 829-07910 )
   
7. Poly(dI-dC) (Sigma-Aldrich, catalog number: 4929 )
   
8. Tris
   
9. Boric acid
   
10. EDTA 2Na
    
11. NaCl
    
12. HCl
    
13. Acrylamide
    
14. N,N’-Methylene-bisacrylamide
    
15. Glycerol
    
16. Triton X-100
    
17. Phenylmethylsulfonyl fluoride (PMSF)
    
18. β-mercaptoethanol
    
19. Ammonium persulfate (APS)
    
20. N,N,N',N'-tetramethylethylenediamine (TEMED)
    
21. 10x TBE( see Recipes)
    
22. 4% native polyacrylamide gel (see Recipes)
    
23. Native-PAGE running buffer (see Recipes)
    
24. Protein storage buffer (see Recipes)
    

Equipment

1. Odyssey CLx Infrared Imaging System (LI-COR)
   
2. A set of devices for polyacrylamide gel electrophoresis
   
3. Power supply
   
4. Refrigerator or cold room
   
5. Heat block
   

Procedure