4-Methylumbelliferyl-β-D-cellobioside acts as a substrate used to measure the activity of (1,4)-β-Glucanase.

(1,4)-β-Glucanase is an enzyme that breaks down glucose by disrupting the glycosidic bonds. This enzyme is also involved in the cell wall synthesis for various fungi.

4-Methylumbelliferyl-β-D-cellobioside can also be used in Cel D Fusion screenings and activity staining in polyacrylamide gels, yielding blue fluorescence as a result.