The Ti-4Al-2V (wt. %) titanium alloy has garnered widespread applications across diverse fields due to its exceptional strength-to-weight ratio, high toughness, specific strength, and corrosion resistance. The welding of Ti-4Al-2V titanium alloy components is often necessary in manufacturing processes, where the reliability of a welded joint critically influences the overall service life of these components. Consequently, a comprehensive understanding of the welded joint’s microstructure and mechanical properties is imperative. In this study, Ti-4Al-2V titanium alloy was welded using multi-layer and multi-pass TIG welding techniques, and a detailed examination was conducted to analyze the microstructure and grain morphology of each microzone of the welded joint. The results revealed the presence of an initial α phase and a secondary lamellar α phase in the heat affected zone (HAZ). Meanwhile, the fusion zone (FZ) primarily comprised a coarse secondary α phase and a small amount of an acicular martensitic α’ phase. Both the recrystallization zone and the superheated zone exhibited a distinct preferred orientation, with grains smaller than 10 μm accounting for 65.9% and 55.1%, respectively. To assess the mechanical properties of the various microzones and the typical microstructure within the welded joint, nanoindentation tests were performed. The results indicated that the recrystallization zone possessed a higher nanohardness (3.753 GPa) than the incomplete recrystallization zone (3.563 GPa) and the superheated zone (3.48 GPa). Among all the microzones, the FZ exhibited the lowest average nanohardness (3.058 GPa). Notably, the basket-weave microstructure demonstrated the highest average nanohardness, reaching 3.93 GPa. This was followed by the fine-grain microstructure, which possessed a slightly lower nanohardness. The Widmanstätten microstructure, on the other hand, exhibited the lowest nanohardness among the three microstructures within the HAZ. Therefore, the basket-weave microstructure stands out as the most desirable microstructure to achieve in the welded joint. In summary, this study provides a comprehensive characterization and analysis of the microstructure and properties of Ti-4Al-2V titanium alloy TIG welds, aiming to contribute to the optimization of the TIG welding process for Ti-4Al-2V titanium alloy.