This work reported the experimental data of densities, speed of sound, and refractive index of a binary mixture containing ethyl isobutyrate and cyclohexane at different temperatures (293.15, 298.15, 303.15, 313.15, and 323.15 K) and atmospheric pressure. The excess molar volumes, partial molar volumes, coefficient of thermal expansion, deviation of isentropic compressibilities, partial molar compressibilities, and refractive index deviation were calculated. The Redlich−Kister equation was used to correlate excess molar volumes, deviation of isentropic compressibilities, and refractive index deviation. The densities and intermolecular interaction of the binary system were determined by molecular dynamics simulation. The results showed that the theoretical densities of the binary system were consistent with the experimental results. The interaction between ethyl isobutyrate and cyclohexane was van der Waals force, and the main strength was the interaction between oxygen atom in ethyl isobutyrate and hydrogen atom in cyclohexane.