We report evidence of the apical localization of the two Na, K‐ATPase β‐subunit isoforms in cells of the inner ear and of the choroid plexus of the rat. To this end, we generated isoform‐specific antisera against the human Na, K‐ATPase β1 and β2 subunits. These polyclonal rabbit antisera were raised against truncated β‐isoform proteins that were made in E coli with pET expression vectors. Deglycosylation of the native antigen with N‐endoglycosidase F shows four bands in the β1 isoform and five bands in the β2 iso‐form immunoblots. In E15 rat embryos, the β1 isoform was detected in brain, heart and kidney and the β2 isoform only in brain. While β‐subunit mRNA expression (Watts AG, Sanchéz‐Watts G, Emanuel JR, Levenson R 1991 Proc Natl Acad Sci USA 88, 7425–7429), and immunoblotting and enzymatic activity have been determined (Zlokovic BV, Mackic JB, Wang L, McComb JG, McDonough A 1993 J Biol Chem 268, 8019–8025), very little is known about the specific localization of each β‐isoform in the epithelia of choroid plexus and inner ear. Immunocytochemical preparations of 15‐day‐old whole rat embryos and adult rat brain showed an enhanced staining for the β1 and β2 isoforms in the apical membrane of the ampullary crests of the inner ear's semicircular ducts and in the cuboidal cells of the choroid plexus