This paper presents the fragility assessment of non-seismically designed steel moment frames with masonry infills. The assessment considered the effects of multiple earthquakes on the damage accumulation of steel frames, which is an essential part of modern performance-based earthquake engineering. Effects of aftershocks are particularly important when examining damaged buildings and making post-quake decisions, such as tagging and retrofit strategy. The procedure proposed in the present work includes two phase assessment, which is based on incremental dynamic analyses of two refined numerical models of the case-study steel frame, i.e. with and without masonry infills, and utilises mainshock-aftershock sequences of natural earthquake records. The first phase focuses on the undamaged structure subjected to single and multiple earthquakes; the effects of masonry infills on the seismic vulnerability of the steel frame were also considered. In the second phase, aftershock fragility curves were derived to investigate the seismic vulnerability of infilled steel frames with post-mainshock damage caused by mainshocks. Comparative analyses were conducted among the mainshock-damaged structures considering three post-mainshock damage levels, including no damage. The impact of aftershocks was then discussed for each mainshock-damage level in terms of the breakpoint that marks the onset of exceeding post-mainshock damage level, as well as the probability of exceeding of superior damage level due to more significant aftershocks. The evaluation of the efficiency of commonly used intensity measures of aftershocks was also carried out as part of the second phase of assessment.