The laser welding technology of ultra-thick plates has been widely applied in many crucial engineering structures, where its safety and stability still suffer from the residual stress brought by the welding temperature. In this work, experimental and numerical studies are carried out to reveal the evolution and distribution of the residual stress. A 5A06 aluminum alloy plate with 30 mm thickness is used to conduct the laser welding experiment, and the short-wavelength X-ray diffractometer (SWXRD) is employed to measure the residual stress. Furthermore, a multi-pass laser welding numerical model is developed to predict the residual stress during the welding process. The simulation results of the weld pool geometry and residual stress agree well with the experimental data, which gives a good validation of the numerical model. Based on the proposed numerical model, the influence of the welding parameters, including inter-pass cooling time and welding rate are also investigated. It is discovered that the reduction of the inter-pass cooling time results in weakness effect of the temperature gradient, which hinders the generation of plastic deformation and leads to a lower residual stress value. Meanwhile, the simulation results also show that the increase in welding rate promotes the generation of thermal stress, which brings a higher residual stress value appeared in the structure.