American Society of Hematology

Figure 5.

$Figure 5. Inhibitory activity of VWF-CP activity of rADAMTS-13 by autoantibodies purified from plasma of patients with acquired TTP. Three IgG fractions were separately purified from 3 patients with acquired (idiopathic) TTP. Preincubated conditioned medium of recombinant wild-type ADAMTS-13 with 10 mM BaCl2 was mixed with an equal volume of purified patient A-C IgGs, (lanes 1-3), healthy IgG (lane 4), or buffer (lane 5), respectively. Then these mixtures were incubated with an equal volume of purified VWF followed by overnight dialysis against 5 mM Tris-HCl, pH 8.0, containing 1.5 M urea at 37°C. An aliquot of each reaction mixture was subjected to 5% SDS-PAGE. The gel was stained with Coomassie Brilliant Blue R-250. (A) Under reducing conditions; (B) under nonreducing conditions.$

Inhibitory activity of VWF-CP activity of rADAMTS-13 by autoantibodies purified from plasma of patients with acquired TTP. Three IgG fractions were separately purified from 3 patients with acquired (idiopathic) TTP. Preincubated conditioned medium of recombinant wild-type ADAMTS-13 with 10 mM BaCl₂ was mixed with an equal volume of purified patient A-C IgGs, (lanes 1-3), healthy IgG (lane 4), or buffer (lane 5), respectively. Then these mixtures were incubated with an equal volume of purified VWF followed by overnight dialysis against 5 mM Tris-HCl, pH 8.0, containing 1.5 M urea at 37°C. An aliquot of each reaction mixture was subjected to 5% SDS-PAGE. The gel was stained with Coomassie Brilliant Blue R-250. (A) Under reducing conditions; (B) under nonreducing conditions.

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