Aging is associated with reduced muscle mass, weakness, and increased fatigability. In skeletal muscle, the Na+-K+ pump (NKA) is important in regulating Na+-K+ gradients, membrane excitability, and thus contractility, but the effects of aging on muscle NKA are unclear. We investigated whether aging is linked with reduced muscle NKA by contrasting muscle NKA isoform gene expression and protein abundance, and NKA total content in 17 Elderly (66.8 ± 6.4 yr, mean ± SD) and 16 Young adults (23.9 ± 2.2 yr). Participants underwent peak oxygen consumption assessment and a vastus lateralis muscle biopsy, which was analyzed for NKA α1-, α2-, α3-, β1-, β2-, and β3-isoform gene expression (real-time RT-PCR), protein abundance (immunoblotting), and NKA total content ([3H]ouabain binding sites). The Elderly had lower peak oxygen consumption (−36.7%, P = 0.000), strength (−36.3%, P = 0.001), NKA α2- (−24.4%, 11.9 ± 4.4 vs. 9.0 ± 2.7 arbitrary units, P = 0.049), and NKA β3-protein abundance (−23.0%, P = 0.041) than Young. The β3-mRNA was higher in Elderly compared with Young ( P = 0.011). No differences were observed between groups for other NKA isoform mRNA or protein abundance, or for [3H]ouabain binding site content. Thus skeletal muscle in elderly individuals was characterized by decreased NKA α2- and β3-protein abundance, but unchanged α1 abundance and [3H]ouabain binding. The latter was likely caused by reduced α2 abundance with aging, preventing an otherwise higher [3H]ouabain binding that might occur with a greater membrane density in smaller muscle fibers. Further study is required to verify reduced muscle NKA α2 with aging and possible contributions to impaired exercise capability and daily living activities.