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. Author manuscript; available in PMC: 2009 Aug 4.
Published in final edited form as: Nat Protoc. 2009;4(3):341–355. doi: 10.1038/nprot.2008.250

Figure 6.

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.