Humans are unique among all species in their ability to develop and enforce social norms, but there is wide variation in the strength of social norms across human societies. Despite this fundamental aspect of human nature, there has been surprisingly little research on how social norm violations are detected at the neurobiological level. Building on the emerging field of cultural neuroscience, we combine noninvasive electroencephalography (EEG) with a new social norm violation paradigm to examine the neural mechanisms underlying the detection of norm violations and how they vary across cultures. EEG recordings from Chinese and US participants (n = 50) showed consistent negative deflection of event-related potential around 400 ms (N400) over the central and parietal regions that served as a culture-general neural marker of detecting norm violations. The N400 at the frontal and temporal regions, however, was only observed among Chinese but not US participants, illustrating culture-specific neural substrates of the detection of norm violations. Further, the frontal N400 predicted a variety of behavioral and attitudinal measurements related to the strength of social norms that have been found at the national and state levels, including higher culture superiority and self-control but lower creativity. There were no cultural differences in the N400 induced by semantic violation, suggesting a unique cultural influence on social norm violation detection. In all, these findings provided the first evidence, to our knowledge, for the neurobiological foundations of social norm violation detection and its variation across cultures.culture | social norms | N400 | electroencephalography | EEG