Painful events are suggested to elicit two opponent responses: a negatively valenced and a positively valenced process triggered by their onset and termination, respectively. Consequently, stimuli (conditioned stimulus, CS) associated with pain onset (unconditioned stimulus, US) provoke defensive responses like startle potentiation, while stimuli associated with pain termination elicit appetitive responses like startle attenuation. Here we summarize four studies elucidating the time course of the relief following the termination of a painful US. In these studies, the timing of a relief-associated stimulus (reliefCS) presented after a painful US was varied from immediately (Study1), 3 seconds (Study4), or 6 seconds (Study2 and 3) after the US. Responses to these relief CSs were compared also with responses to a stimulus presented before the US (fearCS), and a third stimulus (safetyCS) that was never associated with the US. The synopsis of these studies reveals that the timing of the reliefCS is crucial to turn a defensive response into an appetitive response. Namely, the reliefCS immediately following the US-induced startle potentiation and negative valence (Study1); 3 seconds later, the startle response was slightly less potentiated and the ratings a little less negative (Study4); finally, the reliefCS presented 6 seconds after the US caused startle attenuation (Study2 and 3) and positively valenced ratings (Study3). In sum, the observed time course of relief closely follows the predictions of opponent process theory. This means that relief may be a reinforcer that elicits conditioned appetitive behavior, but its reinforcing properties strongly depend on its temporal relation to the aversive event.