GENERATION AND CHARACTERIZATION OF SUBTYPE-SPECIFIC MONOCLONAL-ANTIBODIES TO K-SUBUNIT AND BETA-SUBUNIT POLYPEPTIDES( CHANNEL ALPHA)

Molecular characterization of mammalian voltage-sensitive K+ channel g enes and their expression became possible with the cloning of the Shak er locus of Drosophila. However, analysis of the expression patterns a nd subunit composition of native K+ channel protein complexes requires immunological probes specific for the individual K+ channel gene prod ucts expressed in excitable tissue. Here, we describe the generation a nd characterization of monoclonal antibodies (mAbs) against eight dist inct mammalian K+ channel polypeptides; the Kv1.1, Kv1.2, Kv1.4, Kv1.5 and Kv1.6 Shaker-related alpha-subunits, the Kv2.1 Shab-related alpha -subunit, and the Kv beta 1 and Kv beta 2 beta-subunits. We characteri zed the subtype-specificity of these mAbs against native K+ channels i n mammalian brain and against recombinant K+ channels expressed in tra nsfected mammalian cells. In addition, we used these mAbs to investiga te the cellular and subcellular distribution of the corresponding poly peptides in rat cerebral cortex, as well as their expression levels ac ross brain regions. Copyright (C) 1996 Elsevier Science Ltd