Ischemic preconditioning has been shown to confer ergogenic benefit to aerobic exercise performances, but whether or not this is conferred to the muscle directly via a blood-borne component remains unknown. We tested the possibility that serum from humans following muscle contraction, ischemic preconditioning, or both could affect contractile properties of mouse muscle. No differences in indices of contractile function, fatigue, or recovery were seen between conditions. Further, no human participants improved performance in a 4-kilometre cycling time trial in response to ischemic preconditioning performed with or without exercise compared to control or free flow exercise conditions. Our findings indicate that the humoral component of ischemic preconditioning does not impart ergogenic benefit at the cellular level, that ischemic preconditioning may not manifest in performance benefits unless at high intensities, and that augmented ischemic preconditioning may have a hormetic relationship with performance improvements.