Materials and Reagents

1. Purified ISKNV (was purified in our laboratory)
   
2. 12% SDS-PAGE gel
   
3. PVDF membrane (Whatman, catalog number: 10485290 )
   
4. Coomassie brilliant blue G-250 (Beyotime Institute of Biotechnology, catalog number: P0017B )
   
5. Tris-HCl (pH 7.4)
   
6. EDTA
   
7. Tween 20
   
8. Guanidine hydrochloride (Sigma-Aldrich, catalog number: G4505 )
   
9. Nonfat milk powder
   
10. Maltose-binding protein (MBP) or MBP-mCav-1 protein (these proteins were made in our laboratory)
    
11. Anti-MBP antibodies (Sigma-Aldrich, catalog number: M1321 )
    
12. Horse radish peroxidase (HRP)-conjugated goat anti-mouse secondary antibodies (Invitrogen, catalog number: 626520 )
    
13. HRP substrate solution (Beyotime Institute of Biotechnology, catalog number: P0209 )
    
14. BCA protein assay kit (Thermo Fisher Scientific, catalog number: 23225 )
    
15. Renaturing buffer (see Recipes)
    
16. Tris Buffered Saline and Tween 20 (TBST) (see Recipes)
    

Equipment

1. Electrophoresis equipment
   
2. Bio-Rad blotting equipment (Bio-Rad Laboratories, model: A101441 )
   

Procedure