Imubind tissue factor elisa

After collection, BALFs were immediately put on ice and then centrifuged for 10 min at 900 ×g at 4 °C. The supernatant was aliquoted and stored at − 80 °C until assays were performed. The levels of prothrombin fragment 1 + 2 (F_{1 + 2}), thrombin–antithrombin complex (TATc), soluble TF (sTF) antigen, plasminogen activator inhibitor 1 (PAI‐1) and tissue‐type plasminogen activator (t‐PA) were measured with specific enzyme immunoassays, according to the manufacturers’ instructions (F_{1 + 2}, Enzygnost F_{1 + 2} [Siemens, Marburg, Germany]; TATc, Enzygnost TAT micro [Siemens]; sTF, Imubind Tissue Factor ELISA [Sekisui Diagnostics, Stamford, CT, USA]; PAI‐1 activity, Technozym PAI‐1 Actibind ELISA [Technoclone, Vienna, Austria]; PAI‐1 antigen, Quantikine Human Total Serpin E1/PAI‐1 Immunoassay [R&D Systems, Minneapolis, MN, USA]; and t‐PA antigen and activity, Technozym t‐PA Combi Actibind ELISA [Technoclone]). The F_{1 + 2}, TATc and sTF assays are sensitive to detect even low values in plasma and BALF of healthy volunteers as previously described 22, 23, 24. The lower limits of quantification were 20 pmol L⁻¹ for F_{1 + 2}, 2 μg L⁻¹ for TATc, 0.05 ng mL⁻¹ for sTF, 0.49 IU mL⁻¹ for PAI‐1 activity, 0.313 ng mL⁻¹ for PAI‐1 antigen, 0.05 IU mL⁻¹ for t‐PA activity and 0.1 ng mL⁻¹ for t‐PA antigen). Fibrinogen was measured with the Clauss method in an accredited routine laboratory.