There is an indispensable need for force calibration for quantitative nanoscale force measurement using atomic force microscopy. Calibrating the normal force is relatively straightforward, whereas doing so for the lateral force is often complicated because of the difficulty in determining the optical lever sensitivity. In particular, the lateral force calibration of a colloidal probe in a liquid environment often has a larger uncertainty as a result of the effects of the epoxy, the location of the colloidal particle on the cantilever, and a decrease in the quality factor. In this work, the lateral force of a colloidal probe using a reference cantilever with a known spring constant was calibrated in a liquid environment. By obtaining the spring constant and the lateral sensitivity at the equator of a spherical colloidal particle, the damage to the bottom surface of the colloidal particle could be eliminated. Further, it was shown that the effect of the contact stiffness on the determination of the lateral spring constant of the cantilever could be minimized. It was concluded that this method can be effectively used for the lateral force calibration of a colloidal probe in a liquid environment.