Ab234559

We used multiple assays to comprehensively evaluate VWF, including: 1) VWF antigen (VWF:Ag) and VWF propeptide (VWF:pp) measured by enzyme-linked immunosorbent assay (Cat #: ab223864 [Abcam] and CS-MW1939 [Cell Sciences]); 2) thiols exposed on the surface of VWF multimers (Supplemental Methods); 3) ristocetin-induced platelet agglutination (RIPA) (Helena Laboratories); 4) VWF binding to collagen (VWF:CB) (Supplemental Methods); 5) platelet adhesion under flow conditions (Supplemental Methods); 6) VWF multimers using nonreducing sodium dodecyl sulfate–agarose gel electrophoresis (Supplemental Methods);²⁷ (link) 7) shear-induced platelet aggregation (SIPA) (Supplemental Methods); 8) the exposure of the A1 domain on VWF using immunoprecipitation (Supplemental Methods); 9) endothelial permeability (Supplemental Methods); and 10) ADAMTS-13 antigen (ADAMTS-13:Ag) using enzyme-linked immunosorbent assay (Cat #: ab234559; Abcam) and cleaving an exogenous VWF peptide measured using a FRET (fluorescence resonance energy transfer) assay (Diapharma).

All routine lab tests were performed at the hospital’s clinical chemistry department. ELISA’s were used to measure the plasma protein concentrations for ANGPT2 (DANG20, R&D Systems, Minneapolis, MN, USA), ANGPT1 (DANG10, R&D Systems, Minneapolis, MN, USA), von Willebrand factor (ab108918, Abcam, Cambridge, UK), and ADAMTS13 (ab234559, Abcam, Cambridge, UK), according to the manufacturer’s instructions. Data for thrombin–antithrombin (TAT) complexes were available for some patients from a previous study [10 (link)].