The rejection of unfair offers in the ultimatum game (UG) indicates negative reciprocity. The model of strong reciprocity claims that negative reciprocity reflects prosociality because the rejecting individual is sacrificing resources in order to punish unfair behavior. However, a recent study found that the rejection rate of unfair offers is linked to assertiveness (status defense model). To pursue the question what drives negative reciprocity, the present study investigated individual differences in the rejection of unfair offers along with their behavioral and neuronal determinants. We measured fairness preferences and event-related potentials (ERP) in 200 healthy participants playing a computerized version of the UG with pictures of unfair and fair proposers. Structural equation modeling (SEM) on the behavioral data corroborated both the strong reciprocity and the status defense models of human cooperation: Not only more prosocial but also more assertive individuals were more likely to show negative reciprocity by rejecting unfair offers. Experimental ERP results confirmed the feedback negativity (FN) as a neural signature of fairness processing. Multilevel SEM of brain–behavior relationships revealed that negative reciprocity was significantly associated with individual differences in FN amplitudes in response to proposers. Our results confirm stable individual differences in fairness processing at the behavioral and neuronal level.