Fitc conjugated anti human pac 1

20(S)-protopanaxadiol (PPD, purity of ≥98% by HPLC) was purchased from Shanghai yuanye Bio-Technology Co., Ltd (Shanghai, China). Hemocoagulase was purchased from Jinzhou Ahon Pharmaceutical Co, Ltd. (Liaoning, China). Prothrombin (PT), activated partial thromboplastin time (APTT), thrombin time (TT), and fibrinogen (FIB) kits were obtained from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Luciferin/luciferase reagent and thrombin were purchased from Chrono-Log Corporation (Pennsylvania, USA). Vorapaxar, ticagrelor, and seratrodast were purchased from MedChemExpress (New Jersey, USA). Cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), and human protease activated receptor 1 ELISA kits were purchased from Sino Best Biological Technology Co., Ltd. (Shanghai, China). FITC-conjugated anti-human CD62P (P-selectin) and FITC-conjugated anti-human PAC-1 were purchased from BioLegend (California, USA). Fluo-3 AM calcium indicators and lactate dehydrogenase (LDH) cytotoxicity assay kits were purchased from Beyotime Biotechnology (Shanghai, China).

Platelet agonist ADP and collagen (equine tendon) were from HELENA laboratories (USA). Ristocetin was from Sigma (R7752, USA). Anti-human GPIbα monoclonal antibody SZ2, VM16d, and AK2 were from GenTex (GTX28822, USA), YO Proteins (656, USA), and Bio-Rad (MCA740T, USA), respectively. Secondary antibody anti-human/mouse CD62P (P-selectin) APC was from Thermo Fisher scientific (17-0626, USA), and FITC-conjugated anti-human PAC-1 was from Biolegend (362803, USA). Peptides of GPIbα fragments were synthesized by GL Biochen (China) Ltd. Recombinant mouse vWF protein was from Creative BioMart (VWF-1432 M, USA), and human vWF protein was from Sino Biological (10973-H08C, USA). C57BL/6J mice, BCLB/C mice, and Wistar rat were from JSJ laboratories (China) and were bred and housed at Putuo animal care facility. Transgenic mice expressing no mouse but only human GPIbα (hTg) were described previously [19 (link)]. Animal experiments were conducted in mice or rats using protocols approved by the IACUC of Putuo Hospital. Six- to eight-week-old mice or rats were used in the study, and investigators were blinded to group allocation during data collection.