Evaluation of a new mouse WB-TG assay. (A) Citrated mouse WB was triggered by 0.5 pM TF and 16.7 mM CaCl2, in presence of 416.7 μM thrombin substrate ZGGR-AMC. Cleavage of the thrombin substrate was monitored with Ex = 355 nm and Em = 460 nm. The thrombin-like amidolytic activity was calculated from the first derivative of the fluorescence signal of TG reactions and calibration reactions. WB-TG curves of 6 replicates are shown. (B) The effect of predilution of mouse WB samples on WB-TG was tested by comparing 0.5 pM TF-triggered TG reactions in WB samples that were not prediluted (30 μL WB in 60 μL reaction mixture) against those prediluted (24, 15, or 9 μL WB in 60 μL mixture, indicated as 24/30 prediluted, 15/30 prediluted, and 9/30 prediluted, respectively). Averaged curves of 3 independent experiments are shown. (C-D) Mouse WB-TG in response to different concentrations of TF or silica. Representative curves of 3 independent experiments are shown.
