Background: Only few studies on gender-related differences in skin physiology have been performed – and they provided conflicting results. Objective: In the present study, the effect of gender on the physiology of the stratum corneum was studied. Methods: The physiological parameters transepidermal water loss, pH value, hydration and sebum content were determined on the flexor forearms of 6 female and 6 male volunteers. In addition, stratum corneum samples, removed by tape stripping, were studied for amount, spectroscopic properties, protein content, and mass. Results: The skin of women was characterized by a significantly higher pH value (5.6 ± 0.4) than that of men (4.3 ± 0.4, p < 0.05). Protein absorption was the only other parameter significantly dependent on gender (p < 0.05). Both effects might be caused by differences in human biology, such as hormonal status. Conclusion: Therefore, the gender of the volunteers should be considered in dermatologic studies.