Spot welding which use the main process for side block production of stainless steel railway vehicle is legged behind in laser welding about a quality and productivity. Although the laser welding has many potential advantages such as low heat input and aspect ratio of weld bead, its application to a new structural component still is required many engineering data including mechanical properties such as tensile, fatigue strength, etc on. Therefore, experimental analysis was carried out to understand the fatigue phenomena of different thickness stainless steel overlap joining panels by Nd:YAG laser welding. The fatigue life curves were obtained through fatigue tests with the various levels of applied load. The fatigue life is related with the parameters such as gap size and penetration depth through experiment. As the results, tensile strength and fatigue life were proportional in heat input level and gap was identified the major factor for fatigue life. Also we could know that deferent a-ferrite content at HAZ depend on welding heat input was important factor to determine a formation of initial crack and total fatigue life cycle.