[Production of specific antibodies against isoforms of the beta subunit of human Na,K-ATPase]

The study of the differential and functional tissular distribution of the isoforms of Na,K-ATPase, an enzyme located on the plasma membrane of animal cells, must be performed with the aid of antibodies capable of specifically recognising beta 1 and beta 2 subunits. In order to facilitate the study of the functions and distribution of such isoforms, proteins were generated using pET series plasmid construction and transformation in E coli bacteria, and their production was promoted with IPTG, these being the antigens that induce antibodies specific for each isoform. The antisera thus obtained were used in Western Blot assays with tissue microsomes from different animal species; specifically, beta 1 and beta 2 isoforms of Na,K-ATPase could be recognised. A panel of polyclonal antibodies was obtained which proved capable of recognising these isoforms in different animal species, as well as their deglycosylated isoform. Such antibodies can be used as an efficient tool for functional and tissue distribution studies of the two isoforms.