To determine the accuracy of palpating the long head of the biceps tendon (LHBT) within the intertubercular groove with the use of ultrasonographic localization as a gold standard. Prospective, single-blinded pilot study. Sports medicine clinic at a tertiary care academic institution. Twenty-five male and female asymptomatic volunteers ages 24-41 years (mean, 30.9 ± 4.3 years) with body mass indices of 19.3 to 36.3 kg/m(2) (23.84 ± 4.8 kg/m(2)). Three examiners of differing experience (a sports medicine board-certified staff physician, a sports medicine fellow, and a physical medicine and rehabilitation resident) identified the LHBT location in the intertubercular groove via palpation on a subject in the supine position and marked its location by taping an 18-gauge Tuohy needle to the skin overlying the groove. The examiner order was randomized. A fourth examiner who was blinded to the palpation order assessed the previous examiner's palpation accuracy by comparing the needle position to the sonographically determined tendon position. Needle placement in relation to the intertubercular groove was graded as being within the groove, medial to the groove, or lateral to the groove. In the latter 2 cases, the distance from the needle to the closest groove edge was recorded. Overall accuracy rate was 5.3% (4/75), ranging from 0% (0/25) for the resident to 12% (3/25) for the fellow (P ≤ .007 for interexaminer differences). All missed palpations were localized medial to the intertubercular groove by an average of 1.4 ± 0.5 cm (range, 0.3 for the fellow to 3.5 cm for the resident). Based on the current methodology, clinicians have a tendency to localize the intertubercular groove medial to its actual location. Consequently, clinicians should exercise caution when relying on clinical palpation to either diagnose a biceps tendon disorder or perform a bicipital tendon sheath injection. When clinically indicated, sonographic guidance can be used to accurately identify the LBHT within the intertubercular groove.