This study explored the roles of self-soothing ability and emotional dysregulation in the relationship between second separation-individuation, binge-eating, and self-harm behavior. Using a sample of 238 college students, we studied the relationships among second separation-individuation, self-soothing ability, emotional dysregulation, binge-eating, and self-harm behavior. The results showed that second separation-individuation was positively correlated with self-soothing ability, and negatively correlated with emotional dysregulation, binge-eating, and self-harm behaviors. Furthermore, self-soothing ability and emotional dysregulation mediated the effect of second separation-individuation on binge-eating sequentially. Self-soothing ability, not emotional dysregulation, mediated the effect of second separation-individuation on self-harm behavior, However, in a sample of 89 participants with a history of self-harm, the sequential mediating effect of self-soothing ability and emotional dysregulation in the relationship between second separation-individuation and self-harm behavior was significant. The implications and limitations of this study and future directions of research are discussed.