Quantitative immunoassay of total cellular gap junction protein connexin32 during liver regeneration using antibodies specific to the cooh-terminus

E. Michael Hendrix, Carol Skowron Lomneth, William W. Wilfinger, Elliot L. Hertzberg, Simon J.T. Mao, Lin Chen, William J. Larsen

Research output: Contribution to journalArticle

12 Scopus citations

Abstract

A radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA) were used to determine relative concentrations of liver connexin32 (CX32) in rats. The RIA and ELISA utilize synthetic peptides corresponding to regions of the carboxyl-terminus and antibodies raised in rabbits against these peptides. Assuming that affinities of antisera are similar for peptide and native CX32, total cellular CX32 was found to exceed the amount of gap junction protein at the cell surface calculated from morphometric analyses by 1.5-2.0 fold. This finding raises the possibility that some of the protein is present in cy toplasmic compartments or as occult precursors in the plasma membrane. Studies of CX32 content in regenerating rat liver support this conclusion and show a time course of loss and recovery of CX32 that agrees with those reported in studies using other techniques.

Original languageEnglish (US)
Pages (from-to)61-73
Number of pages13
JournalTissue and Cell
Volume24
Issue number1
DOIs
StatePublished - 1992
Externally publishedYes

Keywords

  • ELISA
  • Liver
  • connexin32
  • radioimmunoassay
  • site specific antibodies

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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