This study investigates the microstructure and nano-hardness distribution across the thickness of an H-section steel beam, specifically designed for seismic and fire resistance and fabricated using a quenching and self-tempering process. The beam dimensions include a 24 mm thick flange, with flange and web lengths of 300 mm and 700 mm, respectively. Our findings indicate that the mechanical properties across the flange thickness meet the designed criteria, with yield strengths exceeding 420 MPa, tensile strengths of over 520 MPa, and a yield-to-tensile strength ratio below 0.75. Microstructurally, the central part of the flange predominantly consists of granular bainite with a small fraction of martensite–austenite (MA) constituents, while locations closer to the surface show increased acicular ferrite and decreased MA constituents due to faster cooling rates. Furthermore, thermal exposure at 600 °C reveals that while the matrix microstructure remains thermally stable, the MA phase undergoes tempering, leading to a decrease in nano-hardness. These insights underline the significant impact of MA constituents on the elongation properties and stress concentrations, contributing to the overall understanding of the material’s behavior under seismic and fire conditions. The study’s findings are crucial for enhancing the reliability and safety of construction materials in demanding environments.