4-Methylumbelliferyl β-D-Glucopyranoside

4-Methylumbelliferyl β-D-Glucopyranoside, a β-D-glucoside, is a fluorogenic substrate for β-glucosidase, utilizes to assay β-glucosidase activity^[1]. 4-Methylumbelliferyl β-D-Glucopyranoside releases the highly fluorescent 4-methylumbelliferyl (4-MU), which has an emission maximum at 445-454 nm. The excitation maximum for 4-MU is pH-dependent: 330, 370, and 385 nm at pH 4.6, 7.4, and 10.4, respectively^[2]. IC50 & Target:β-glucosidase^[1] In Vitro:The following are the experimental steps for the β-glucosidase GLB1 luciferase activity assay using 4-Methylumbelliferyl β-D-glucopyranoside as a substrate:
1. Reagent preparation
0.64 mM 4-Methylumbelliferyl β-D-glucopyranoside, 50 mM sodium citrate (pH 3.5), recombinant human GLB1 protein expressed in CHO cells.
2. Sample treatment
Incubate at 37°C for 20 minutes and terminate the reaction with alkaline stop solution.
3. Standard curve generation
Generate a standard curve using 0.01-3 nmol/tube of 4-Methylumbelliferone (HY-N0187). The results are linear over the enzyme amount range of 10-300 ng/tube.