To optimize the tempering process of 40Cr13VTi steel containing V and Ti, the effect of tempering temperature on microstructure, precipitation, and properties of 40Cr13VTi stainless steel has been studied. Optical microscope (OM), scanning electron microscopy (SEM), and energy dispersive spectrometer (EDS) were used to analyze microstructure and precipitates. Tensile, impact, and hardness testing tests were performed to test the mechanical properties of 40Cr13VTi martensitic stainless steel. The research results indicate that as the increase of tempering temperature, the carbides precipitated gradually increase, and the shape gradually changes from particle to flake, which is consistent with the direction of the original martensite flat surface, and most of them concentrate at the martensite boundary. When the tempering temperature is 350°C, 40Cr13VTi martensitic stainless steel obtains the best comprehensive properties, strength, impact toughness (IT), and hardness which are 1360 MPa, 1717 MPa, 8.5 J/cm2, and 45.1 HRC respectively.