We prepared the substrate by shaking one chicken egg yolk in a solution
of 150mM NaCl to a final volume of 100mL and stored this at 4 C
prior to the reaction.In each assay,we prepared several tubes by
taking a final volume(0.33mL)of a10% dilution of the egg yolk
suspension and adding it to a solution containing 150mM NaCl,
10mM CaCl2, 0.01% taurocholic acid,and 5.0mM Tris–HCl (pH
7.4),The tubes were kept at 37 C during the procedure.