“…Analysis of surface glycoproteins (GP) Ibα (by a monoclonal anti‐human CD42b antibody coupled with phycoerythrin [PE], Dako), GPIIb‐IIIa (anti‐hCD41‐fluorescein isothiocyanate [FITC] and anti‐hCD61‐FITC, Becton Dickinson), baseline expression P‐selectin expression (anti‐CD62P‐PE, Becton Dickinson), and PAC1 binding (PAC1‐FITC, Becton Dickinson) were performed in a 100‐µL volume containing cryopreserved PLTs at a final concentration of 5 × 10 9 /L and combinations of relevant antibodies at saturating concentrations. After a 15‐minute incubation at room temperature in the dark, 1000 µL of Tyrode's buffer was added and cryopreserved PLTs were immediately analyzed by flow cytometry (FACSCanto, Becton Dickinson) . Dose response of PLT reactivity was investigated with ADP (baseline and final ADP concentrations of 0.5, 5.0, and 50 µmol/L), convulxin (0.5, 5, and 50 ng/mL), or thrombin (0.005, 0.05, and 0.5 U/mL) in a 100‐µL volume containing cryopreserved PLTs at a concentration of 5× 10 9 /L, anti‐CD62P‐PE, and PAC1‐FITC.…”