Fig. 7.

Simultaneous binding of α₂-antiplasmin, tPA, and plasminogen to immobilized fibrin detected by ELISA. A mixture of 0.5 μM biotinylated α₂-AP, 2.5 μM tPA, and 2.5 μM plasminogen was added to surface-adsorbed fibrin in the absence (filled bars) or presence (empty bars) of 100 mM ε-ACA. Bound biotinylated α₂-AP was detected spectrophotometrically at 405 nm using avidin conjugated to alkaline phosphatase, bound tPA and plasminogen were detected spectrophotometrically at 450 nm using the anti-tPA monoclonal antibody and specific polyclonal antibodies, respectively, as described in Experimental Procedures. All results are means ± the standard deviation of two independent experiments, each performed in duplicate.