Purified human prothrombin was from Synapse Research Institute (Maastricht, The Netherlands) and rivaroxaban (Xarelto) was from Bayer Schering Pharma AG. Anti‐FcγRIIA monoclonal antibody (mAb) IV.3 was purified in the laboratory from a hybridoma.12 (link) The antiprothrombin mAb 28F4 is a mouse IgG antibody, as described previously.13 (link), 14 (link) Antiprothrombin fragment 1+2 mAbs 3B1, 6A3, 11H2, and 8H11 were developed and produced according to standard procedures.15 (link) LAC was measured by activated partial thromboplastin time (Diagnostica Stago), dilute Russell Viper Venom Time (Diagnostica Stago), and Ecarin Clotting Time (ECT; Diagnostic Reagents). The Pierce Classic IP Kit from Thermo Fisher Scientific was used for immunoprecipitation. High‐binding enzyme‐linked immunosorbent assay (ELISA) plates from Costar were used for the ELISAs. Sheep anti‐human prothrombin IgG horseradish peroxidase–labeled antibody was from Affinity Biologicals, Inc., prothrombin fragment 1+2 was from Haematologic Technologies, Inc. All other reagents are previously described or are from Sigma‐Aldrich.16 (link)
Protocol detail
Antiprothrombin Antibody Characterization
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Activated partial thromboplastin time
Anti antibody
Anti igg
Antibody
Antibody iv
Biologicals
Diagnostic
Dilute
Ecarin
Elisas
Fcγriia
Horseradish peroxidase
Human
Hybridoma
Igg antibody
Immunoprecipitation
Mabs
Mouse
Prothrombin
Prothrombin fragment 1 2
Rivaroxaban
Russell viper venom time
Sheep
Synapse
Xarelto
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Variable analysis
independent variables
- Rivaroxaban (Xarelto) from Bayer Schering Pharma AG
- Anti-FcγRIIA monoclonal antibody (mAb) IV.3 purified in the laboratory from a hybridoma
- Antiprothrombin mAb 28F4 (a mouse IgG antibody)
- Antiprothrombin fragment 1+2 mAbs 3B1, 6A3, 11H2, and 8H11 developed and produced according to standard procedures
- LAC measured by activated partial thromboplastin time (Diagnostica Stago), dilute Russell Viper Venom Time (Diagnostica Stago), and Ecarin Clotting Time (ECT; Diagnostic Reagents)
- Purified human prothrombin from Synapse Research Institute (Maastricht, The Netherlands)
- Sheep anti-human prothrombin IgG horseradish peroxidase–labeled antibody from Affinity Biologicals, Inc.
- Prothrombin fragment 1+2 from Haematologic Technologies, Inc.
- All other reagents previously described or from Sigma-Aldrich
- Not explicitly mentioned
- Not explicitly mentioned
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