The Na,K-ATPase comprises a family of isozymes that catalyze the active transport of cytoplasmic Na+ for extracellular K+ at the plasma membrane of cells. Isozyme diversity for the Na,K-ATPase results from the association of different molecular forms of the alpha (alpha1, alpha2, alpha3, and alpha4) and beta (beta1, beta2, and beta3) subunits that constitute the enzyme. The various isozymes are characterized by unique enzymatic properties and a highly regulated pattern of expression that depends on cell type, developmental stage, and hormonal stimulation. The molecular complexity of the Na,K-ATPase goes beyond its alpha and beta isoforms and, in certain tissues, other accessory proteins associate with the enzyme. These small membrane-bound polypeptides, known as the FXYD proteins, modulate the kinetic characteristics of the Na,K-ATPase. The experimental evidence available suggests that the molecular and functional heterogeneity of the Na,K-ATPase is a physiologically relevant event that serves the specialized functions of cells. This article focuses on the functional properties, regulation, and the biological relevance of the Na,K-ATPase isozymes as a mechanism for the tissue-specific control of Na+ and K+ homeostasis.