Electrophoresis and Protein Identification

SDS-PAGE was performed as previously described (Laemmli, 1970). Overnight, 10-17% gradient gels were run with 14 A h Analytical PAA gels were silver-stained according to the method of Heukeshoven and Dernick (1985), and preparative gels were stained with Coomassie brilliant blue. 

For the identification of proteins, preparative gels with a total protein load of 1 mg per gel were mn and proteins were visualized by Coomassie Blue R staining. Leflunomide binding proteins were analyzed using preparative SDS-PAGE. Coomassie-stained protein bands were identified by MS. 

---