Platelet Activation and Signaling Assay

Bovine thrombin, Fluorescein isothiocyanate (FITC), dialysis tubing, wortmannin, TFLLR-NH₂ and AYPGKF-NH₂ were obtained from Sigma (Poole, UK). Bisindolylmaleimide I (Bis-1) and primary 4G10 antibody were purchased from Merck Millipore (Darmstadt, Germany). RWJ56110 and TCY-NH₂ were purchased from R&D Systems (Minneapolis, MN, USA). Primary FITC labeled PAC-1 and Cy5 labelled anti-CD62P were purchased from BD Biosciences (New Jersey, NJ, USA). Staphostatin A was purchased from Biocentrum (Krakow, Poland). Fast flow DEAE and CF sepharose columns were purchased from GE Healthcare (Buckinghamshire, UK). Brain heart infusion media was purchased from Oxoid (Basingstoke, UK). All other reagents were from previously described sources [26 (link)].