Figure 2.

Monitoring of fluorescence by the Endo IV cleavage reaction was performed in 5 mM MgCl₂, 20 mM Tris–HCl (pH 8.5). The three component substrate complex used in these experiments is shown. FL1 is a fluorescein derivative and Q is the Eclipse^® Dark Quencher, structures of which are shown in Figure 3B and D, respectively. The reaction mixture contained Endo IV at 0.04 U/μl concentration, probe and enhancer at 150 nM, and the target at 5 nM. The experiment was performed on LightCycler.