Figure 6.
Western blot analysis showing the IMAC purification steps of a chimeric Flag/MaSp 2 recombinant protein. The 60-kDa His-tagged A4S88 recombinant protein was detected using the 6× His mAb–HRP conjugate. Mk: molecular weight marker Precision Plus Protein Standard Dual color; F1–6: IMAC collected fractions—F1: unbound proteins; F2–4: wash 1 (20 mM imidazole), wash 2 (40 mM imidazole), wash 3 (50 mM imidazole), respectively; F5: elution fraction (250 mM imidazole); F6: strip fraction. The recombinant protein is highly concentrated in the elution fraction (F5) and residual in the strip fraction (F6). The numbers indicate the molecular weights in kDa.