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. Author manuscript; available in PMC: 2020 Feb 1.
Published in final edited form as: Carbohydr Polym. 2018 Oct 28;205:385–391. doi: 10.1016/j.carbpol.2018.10.071

Figure 3.

Figure 3.

Optimization of heparanase assay using heparin-DE as substrate. (A) Labeled heparin (1 mg/ml) was incubated 1 μM HPSE at 37 °C for indicated times followed by measurement of emission at 500 nm. (B) Labeled heparin (1 mg/ml) was incubated with varying concentrations HPSE at 37 °C for 4 h followed by fluorescence measurement. (C) Michaelis–Menten kinetics of HPSE cleavage of heparin–DE using the optimized FRET quenching assay. The cleavage reactions were performed in microplate format (100 μL) in 20 mM sodium acetate buffer, pH 5.0, containing 1 mg/mL heparin–DE and 1 μM HPSE at 37 °C for 4 h. Solid lines represent curve fitting to the standard Michaelis equation. Error bars show variation from at least 3 measurements. F and F0 are fluorescence signals corresponding to the test sample and blank, respectively.