Effects of α2AP and recombinant HSA-related proteins on plasma clot formation and lysis. Clot formation and lysis were followed by monitoring turbidity (absorbance at 340 nm) every 30 seconds for 4 hours using a plate reader, of clots formed using diluted α2AP-deficient plasma, recalcified with 5 mM CaCl2 and supplemented with both 5 nM thrombin and 0.125 nM tPA, and taking the area under the turbidity versus time curve (AUC). Reactions were carried out with or without addition of 1 μM α2AP and/or 20 μM fusion protein. (A) shows results of a single representative experiment. (B) shows quantification of turbidity plots for reactions similar to those shown in A, in the presence or absence of XL5-HSA or XL6-HSA competitor proteins; reaction components are indicated (+ or -) below the graph. The mean ± SD of 5-14 determinations is shown. AUC values were normalized, taking the no tPA, no α2AP condition as 100% (grey bar); white (open) bars correspond to tPA only condition. (C) shows similar results to panel B for additional competitor proteins identified below the graph. As in panel B, the mean ± SD of 5-14 determinations is shown.
Sheffield and Eltringham-Smith BMC Biotechnology 2011 11:127 doi:10.1186/1472-6750-11-127