| Assay Method Information | |
| | β-Gal Inhibitory Assay |
| Description: | β-Gal activity was measured by using 4-methylumbelliferyll-β-D-galactopyranoside in buffer B (0.15 M sodium citrate, pH 4.5, and 0.2 M NaCl) as a substrate, and incubating at 37 °C for 6 min. The reaction was terminated by adding 0.2 M glycine-NaOH buffer (pH 10.7). The liberated 4-methylumbelliferyll was measured with a fluorescence plate reader (excitation 355 nm; emission 460 nm; fluoroskan ascent, Thermo electron Corp.). For inhibition assays, the ligand compounds (galactose, 25 mM; DGJ, 250 μM; NOEV, 2.5 μM; 6S-NBI-DGJ, 250 μM; 6S-NBI-GJ, 2.5 mM; and NBT-DGJ, 2.5mM) were added to the purified β-Gal, and then the activityof β-Gal was measured as described above. |
| Affinity data for this assay | |
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