Generation and characterization of subtype-specific monoclonal antibodies to K+ channel α- and β-subunit polypeptides

Z. Bekele-Arcuri, M. F. Matos, L. Manganas, B. W. Strassle, M. M. Monaghan, K. J. Rhodes, James Trimmer

Research output: Contribution to journalArticle

97 Scopus citations

Abstract

Molecular characterization of mammalian voltage-sensitive K+ channel genes and their expression became possible with the cloning of the Shaker locus of Drosophila. However, analysis of the expression patterns and subunit composition of native K+ channel protein complexes requires immunological probes specific for the individual K+ channel gene products expressed in excitable tissue. Here, we describe the generation and characterization of monoclonal antibodies (mAbs) against eight distinct mammalian K+ channel polypeptides; the Kv1.1, Kv1.2, Kv1.4, Kv1.5 and Kv1.6 Shaker-related α-subunits, the Kv2.1 Shab-related α-subunit, and the Kvβ1 and Kvβ2 β-subunits. We characterized the subtype-specificity of these mAbs against native K+ channels in mammalian brain and against recombinant K+ channels expressed in transfected mammalian cells. In addition, we used these mAbs to investigate the cellular and subcellular distribution of the corresponding polypeptides in rat cerebral cortex, as well as their expression levels across brain regions.

Original languageEnglish (US)
Pages (from-to)851-865
Number of pages15
JournalNeuropharmacology
Volume35
Issue number7
DOIs
StatePublished - 1996
Externally publishedYes

Keywords

  • Cortex
  • Immunoblotting
  • Immunohistochemistry
  • Ion channels
  • Mammalian central nervous system
  • Transfected cells

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Drug Discovery
  • Pharmacology

Fingerprint Dive into the research topics of 'Generation and characterization of subtype-specific monoclonal antibodies to K<sup>+</sup> channel α- and β-subunit polypeptides'. Together they form a unique fingerprint.

  • Cite this

    Bekele-Arcuri, Z., Matos, M. F., Manganas, L., Strassle, B. W., Monaghan, M. M., Rhodes, K. J., & Trimmer, J. (1996). Generation and characterization of subtype-specific monoclonal antibodies to K+ channel α- and β-subunit polypeptides. Neuropharmacology, 35(7), 851-865. https://doi.org/10.1016/0028-3908(96)00128-1