This paper describes the binding of Calcofluor, a fluorescent probe, to oat beta-glucan in buffer solutions. The binding equilibrium constant (K), the total number of binding sites per beta-glucan molecule (N), and the average binding number of Calcofluor per beta-glucan molecule (n) were determined by UV spectroscopic method. The results indicate that the association of Calcofluor and beta-glucan is driven by both enthalpy and entropy and that the process involves hydrogen bonding, van der Waals forces, and hydrophobic interaction. Higher buffer concentration and NaCl facilitate the binding of Calcofluor to beta-glucan. The adsorption isotherm fits a Langmuir model quite well.