Generation and characterization of mouse monoclonal antibodies against CIP75, an UbL-UBA domain-containing protein

Hybridoma (Larchmt). 2009 Apr;28(2):79-84. doi: 10.1089/hyb.2008.0082.

Abstract

CIP75 is a member of the UbL(ubiquitin-like)-UBA (ubiquitin-associated) domain containing protein family, which has a variety of functions. One specific role described for several members of the UbL-UBA family is the involvement in the proteasomal degradation of target proteins. We have reported that CIP75 interacts with the gap junction protein, connexin43 (Cx43), and that CIP75 may modulate the proteasomal degradation of Cx43. Thus, CIP75 may have a critical role in regulating Cx43 levels, and thus intercellular gap junctional communication. This study reports the development of monoclonal antibodies (MAbs) against CIP75 and the characterization of these antibodies through immunoblotting, immunoprecipitation, and immunofluorescence microscopy analyses. These MAbs will be useful tools in future studies to elucidate the role of CIP75 in Cx43 proteasomal degradation as well as other potential activities.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Cloning, Molecular
  • Connexin 43 / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli
  • Genetic Vectors
  • Immunoblotting
  • Immunoprecipitation
  • Mice
  • Microscopy, Fluorescence
  • Proteasome Endopeptidase Complex / metabolism
  • Ubiquitins / immunology*
  • Ubiquitins / metabolism

Substances

  • Antibodies, Monoclonal
  • Connexin 43
  • Ubiquitins
  • Proteasome Endopeptidase Complex