American Society of Hematology

Fig. 6.

$Fig. 6. Gel filtration profile of a prothrombin and patient IgG mixture. / Prothrombin (1 μM) was incubated with 750 μg/mL patient IgG at 4°C for 2 hours. This was followed by the addition of3H-DFP and a further incubation at 25°C for 2 hours. The sample was separated on a Sephacryl S-200HR gel filtration column, and the IgG and prothrombin concentrations of each fraction were measured (A). Arrowhead shows the prothrombin complexed with IgG. Radioactivity in the fractions was determined by counting in a liquid scintillation counter (B). In the absence of DFP treatment, a prothrombin (1 μM) and IgG (750 μg/mL) mixture was applied to the column at 4°C, and each fraction was separately incubated with 5 μM antithrombin III at 37°C for 3 hours. The generation of TAT′ complex was analyzed by ELISA (C).$

Gel filtration profile of a prothrombin and patient IgG mixture.

Prothrombin (1 μM) was incubated with 750 μg/mL patient IgG at 4°C for 2 hours. This was followed by the addition of³H-DFP and a further incubation at 25°C for 2 hours. The sample was separated on a Sephacryl S-200HR gel filtration column, and the IgG and prothrombin concentrations of each fraction were measured (A). Arrowhead shows the prothrombin complexed with IgG. Radioactivity in the fractions was determined by counting in a liquid scintillation counter (B). In the absence of DFP treatment, a prothrombin (1 μM) and IgG (750 μg/mL) mixture was applied to the column at 4°C, and each fraction was separately incubated with 5 μM antithrombin III at 37°C for 3 hours. The generation of TAT′ complex was analyzed by ELISA (C).

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