Figure 1

Protein purification of FCR3 var2CSA DBL domains expressed in HEK293. a. SDS PAGE and Western blot analysis. NuPAGE Novex 4–12% Bis-Tris gels under reduced conditions were loaded with HEK293 extracts obtained after transfection with the different FCR3 var2CSA DBL domains. Gels were either stained with Coomasie blue or transferred to a PVDF membrane to detect recombinant products using an anti-His Tag antibodies. Lanes 1, 2, 3, 4, 5, and 6 correspond respectively to DBL1-X, DBL2-X, DBL3-X, DBL4-ε, DBL5-ε, and DBL6-ε. Culture supernatant, soluble and insoluble fractions after cell lysis are indicated. b. Electrophoresis of purified HEK293 expressed DBL domains. NuPAGE Novex 4–12% Bis-Tris gels under reduced (+DTT) or non reduced (-DTT) conditions were loaded with DBL1-X, DBL3-X, DBL4-ε and DBL6-ε obtained after His-Tag purification and stained with Coomasie blue. Lanes 1, 3, 4, and 6 correspond respectively to DBL1-X, DBL3-X, DBL4-ε and DBL6-ε domains. Protein yields were 0.6 mg/L for DBL1-X, 0.9 mg/l for DBL4-ε and 5 mg/l for DBL6-ε.