Fig. 3.
Fig. 3. Binding of factor IXa to LRP in a solid-phase binding assay in the absence or presence of various components. / Factor IXa (70 nmol/L) was incubated with immobilized LRP (75 ng/well) in a volume of 50 μL in 1% (wt/vol) BSA in HBST in the absence or presence of 1-μmol/L anti–factor IX polyclonal antibodies, 1-μmol/L anti–LRP polyclonal antibodies, and 10-mmol/L EDTA or 0.5-μmol/L urokinase for 1 hour at 37 °C. After washing with HBST, wells were incubated with 2-μg/mL monoclonal anti–factor IX antibody CLB-FIX 14. Bound CLB-FIX 14 was quantified using biotinylated rat antibodies against mouse immunoglobin Gκ chains. Bound complexes were then detected using horseradish peroxidase–labeled streptavidine. Binding is expressed as the percentage of binding in the absence of competitor and is corrected for nonspecific binding (5%-10% relative to binding to LRP coated wells). Data represent the mean and range of 2 independent experiments.

Binding of factor IXa to LRP in a solid-phase binding assay in the absence or presence of various components.

Factor IXa (70 nmol/L) was incubated with immobilized LRP (75 ng/well) in a volume of 50 μL in 1% (wt/vol) BSA in HBST in the absence or presence of 1-μmol/L anti–factor IX polyclonal antibodies, 1-μmol/L anti–LRP polyclonal antibodies, and 10-mmol/L EDTA or 0.5-μmol/L urokinase for 1 hour at 37 °C. After washing with HBST, wells were incubated with 2-μg/mL monoclonal anti–factor IX antibody CLB-FIX 14. Bound CLB-FIX 14 was quantified using biotinylated rat antibodies against mouse immunoglobin Gκ chains. Bound complexes were then detected using horseradish peroxidase–labeled streptavidine. Binding is expressed as the percentage of binding in the absence of competitor and is corrected for nonspecific binding (5%-10% relative to binding to LRP coated wells). Data represent the mean and range of 2 independent experiments.

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