Vacations contribute significantly to employee well-being. However, the positive effects of vacation on employees fade out over time (fade-out effect). Although the phenomenon is robust, little is known regarding the factors that can be expected to preserve employees’ enhanced well-being after returning from vacation. The present study examined organizational variables having the potential to accelerate or delay fade-out effects. Specifically, it was hypothesized that perceived job insecurity and perceived organizational support may affect vacation fade-out effects. A questionnaire was completed by 124 Israeli employees at three time points: before departing for vacation, immediately following the vacation, and 2 weeks after returning from vacation. Findings indicated that employee health significantly increased, and employee exhaustion significantly decreased, after vacation. However, these beneficial effects tended to fade out. Furthermore, upon returning from vacation, perceived organizational support was found to inhibit the increase in employee exhaustion and the decline in health, whereas perceived job insecurity increased employee exhaustion and decreased health. Finally, perceived organizational support buffered the effect of perceived job insecurity on the fade-out effect of exhaustion. Our study contributes to the cumulative research in the field by suggesting that organizational support and job security may be useful variables for exploring the dynamic aspect of employees’ well-being after vacationing.