Integrin αIIbβ3 Binding Assay Protocol

Human integrin αIIbβ3 was purchased from Enzyme Research Laboratories (South Bend, USA). Unless otherwise stated, all mentioned antibodies were bought from Biolegend (San Diego, USA). Dimyristoylphosphatidylglycerol (DMPG; 14:0 PG), dimyristolphosphatidylcholine (DMPC; 14:0 PC) and dioleoyl-glycero-phosphoethanolamine-N-carboxyfluorescein (PE CF) were obtained from Avanti Polar Lipids Inc. (Alabaster, USA). SM-2 biobeads were supplied by Bio-Rad (Munich, Germany). Unfractionated heparin (UFH), fondaparinux and quinine sulfate were purchased from Sigma-Aldrich (Steinheim, Germany). Protein concentration was determined by bicinchonic acid assay (BCA) kit with included protein microstandard (Sigma-Aldrich, Steinheim, Germany). Tris-Base, ethylenediaminetetraacetic acid (EDTA), bovine serum albumin and NaCl were bought from Sigma-Aldrich (Taufkirchen, Germany). CaCl₂, MnCl₂, Triton X-100 and methanol were purchased from Carl Roth GmbH (Karlsruhe, Germany). Sucrose and sodium dodecyl sulfate (SDS) were obtained from Merck KgaA (Darmstadt, Germany).