The purpose of this study was to determine if a difference existed in the rate of fatigue of select shoulder muscles during isometric shoulder elevation and if the measured rate of fatigue was consistent from day to day. Shoulder muscle fatigue has been associated with alterations in joint mechanics and possibly contributes to shoulder dysfunction. While research exists, there is limited information on an objective and reliable measure of shoulder fatigue. Sixteen asymptomatic subjects were evaluated. The subjects held a weight equivalent to 60% of his/her Maximum Voluntary Isometric Contraction (MVIC) while elevating in the scapular plane. Surface electrodes were applied to collect electromyographic activity from the upper trapezius, middle deltoid, serratus anterior, and lower trapezius muscles while the arm was held at 90 degrees elevation. Data collection ceased when the subject was no longer able to maintain 90 degrees of elevation. The subject then rested and a second trial performed. One week later, the two-trial procedure was repeated. A significant interaction of trial x day x muscle was found for the rate of fatigue. Post hoc analysis revealed that the rate of fatigue of the middle deltoid was significantly greater than the other muscles tested. The intraday reliability was good for all muscles but interday reliability was poor except for the middle deltoid. This study suggests that the middle deltoid appears to fatigue faster than the other shoulder muscles tested at the selected level of shoulder elevation. This should be considered in designing a rehabilitation program to develop a sequence that does not overly fatigue the middle deltoid.