American Society of Hematology

Figure 1

$Figure 1. Recombinant Stx1B and Stx2B. Periplasmic extracts (PE) were prepared from Escherichia coli (E coli) BL21(DE3) expressing Stx1B subunit with C-terminal V5 and (His)6 tags (left), or Stx2B subunit with a C-terminal (His)6 tag (right). Extracts were chromatographed on HisPur Cobalt resin (Pierce) and eluted with buffer containing 0.15M imidazole. (A) SDS-PAGE of fractions and standard proteins (Std). StxB subunits were depleted from the flow through (FT) fractions and recovered as pure proteins in the eluate (EL). Purified Stx1B (B) and Stx2B (C) were analyzed TSK G3000SW gel filtration chromatography. The arrows indicate the elution positions, left to right, of thyroglobulin (670 kDa), IgG (158 kDa), ovalbumin (44 kDa), myoglobin (17 kDa), and vitamin B12 (1.35 kDa).$

Recombinant Stx1B and Stx2B. Periplasmic extracts (PE) were prepared from Escherichia coli (E coli) BL21(DE3) expressing Stx1B subunit with C-terminal V5 and (His)₆ tags (left), or Stx2B subunit with a C-terminal (His)₆ tag (right). Extracts were chromatographed on HisPur Cobalt resin (Pierce) and eluted with buffer containing 0.15M imidazole. (A) SDS-PAGE of fractions and standard proteins (Std). StxB subunits were depleted from the flow through (FT) fractions and recovered as pure proteins in the eluate (EL). Purified Stx1B (B) and Stx2B (C) were analyzed TSK G3000SW gel filtration chromatography. The arrows indicate the elution positions, left to right, of thyroglobulin (670 kDa), IgG (158 kDa), ovalbumin (44 kDa), myoglobin (17 kDa), and vitamin B12 (1.35 kDa).

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