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2 protocols using human integrin αiibβ3

1

Integrin αIIbβ3 Binding Assay Protocol

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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).
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2

Integrin αIIbβ3 Reconstitution and Characterization

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Iron(III) nitrate nonahydrate (Fe(NO 3 ) 3 •9H 2 O, >98%), and iron (II) chloride tetrahydrate (FeCl 2 •4H 2 O, 98%) were from Alfa Aesar. Iron(III) chloride hexahydrate (FeCl 3 •6H 2 O, >97%) was obtained from PanReac. Fibrinogen (lyophilized powder) was purchased from Merck KGaA (Darmstadt, Germany) with a purity of 98%. Dextran 6 with a molar mass of 6000 g mol -1 was obtained from Carl Roth GmbH + Co KG (Karlsruhe, Germany). Polyethylene glycol monomethylether phosphonic acid (PEG-PA; 1900 g mol -1 ) was purchased from Sikémia, France.
Human integrin αIIbβ3 was obtained from Enzyme Research Laboratories (South Bend, USA). 1,2-Dimyristoyl-snglycero-3-phosphoglycerol, (DMPG; 14:0 PG) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC; 14:0 PC) were purchased from Avanti Polar Lipids Inc. (Alabaster, USA). SM-2 biobeads were bought from Bio-Rad (Munich, Germany). Tris-Base, ethylenediaminetetraacetic acid (EDTA) and NaCl were supplied by Sigma-Aldrich (Taufkirchen, Germany). CaCl 2 , MnCl 2 , Triton X-100 and methanol were purchased from Carl Roth GmbH (Karlsruhe, Germany). Sucrose, phosphate buffered saline (PBS) and sodium dodecyl sulfate (SDS) were obtained from Merck KGaA (Darmstadt, Germany).
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