American Society of Hematology

Fig. 1.

$Fig. 1. Western blot analysis of immunoprecipitated factor XI secreted by BHK cells transfected with mutant factor XI constructs. / Twenty-microliter samples of WT, mutants, and mock (M) were size-fractionated in 2 experiments (panels A and B) under nonreducing conditions on a 7.5% SDS-PAGE and reacted with polyclonal rabbit antihuman factor XI antibodies. Factor XI dimer was detected only in WT and Tyr493His samples, and factor XI monomer was observed only in the Cys321Phe mutant. A fast-migrating nonspecific band was observed in all samples, including the 2 mocks. A nonspecific slow-migrating band present in panel A most probably corresponds to mouse IgG, which is recognized by the secondary peroxidase-conjugated antibody. This band is absent from samples shown in panel B, probably because the secondary antibody used in this experiment was preadsorbed with mouse IgG, thereby eliminating cross-reactivity.$

Western blot analysis of immunoprecipitated factor XI secreted by BHK cells transfected with mutant factor XI constructs.

Twenty-microliter samples of WT, mutants, and mock (M) were size-fractionated in 2 experiments (panels A and B) under nonreducing conditions on a 7.5% SDS-PAGE and reacted with polyclonal rabbit antihuman factor XI antibodies. Factor XI dimer was detected only in WT and Tyr493His samples, and factor XI monomer was observed only in the Cys321Phe mutant. A fast-migrating nonspecific band was observed in all samples, including the 2 mocks. A nonspecific slow-migrating band present in panel A most probably corresponds to mouse IgG, which is recognized by the secondary peroxidase-conjugated antibody. This band is absent from samples shown in panel B, probably because the secondary antibody used in this experiment was preadsorbed with mouse IgG, thereby eliminating cross-reactivity.

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