| Assay Method Information | |
| | GCase IC50 Assay |
| Description: | The assays were performed at 37°C with 4-MU-β-ᴅ-Glu as the substrate in Mcllvaine buffer (sodium citrate (100mM) and sodium phosphate (200mM); pH 5.2 or 7.0) containing sodium taurocholate (0.25% w/v) and Triton X-100 (0.10% v/v). Enzyme solution (12.5µL, 0.1mg/mL) and inhibitor (7.5µL, various concnetrations) were mixed and incubated at 37°C for 30 min, followed by the addition of substrates (30µL, 4.0mM, in Mcllvaine buffer, pH 5.2 or 7.0) and further incubated for 10 min. The assay was terminated by adding glycine/NaOH buffer (150µL, 100mM, pH 10.6). The amount of released 4-metylumbelliferone (4-MU) was determined by fluorimetry (excitation 355nm, emmision 460nm). |
| Affinity data for this assay | |
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