Nickel-based GH3536 alloy exhibits high strength but low ductility when produced by selective laser melting (SLM), which hinders its widespread application in aerospace. This paper employed a ‘two-step’ method of heat treatment to optimize the microstructure of the SLM GH3536 alloy and investigated the mechanical properties. Tensile tests were conducted on SLM GH3536 alloy before and after heat treatment at 750 °C, using an in-situ high-temperature tensile stage. The relationship between the microstructural evolution and deformation behavior of the alloys was investigated by combining the in-situ tensile experiments and crystal plasticity finite element method (CPFEM). The results suggested that the elongation can be increased by 80% without a significant reduction in tensile strength. The improved elongation is attributed to the complete recrystallization, formation of annealed twins and serrated grain boundaries after heat treatment. The study also investigated the deformation behavior and fracture mode of the alloy before and after heat treatment. Discussion was done on the influence of the microstructure, particularly the twin and serrated grain boundaries on the alloy's plastic deformation. This study enhanced the understanding of plastic deformation of SLM GH3536 alloy and provided a valuable reference for the design and post-treatment of superalloy.