Purification of the insulin-like growth factor II (IGF-II) receptor from an IGF-II-producing cell line, and generation of an antibody which both immunoprecipitates and blocks the type 2 IGF receptor

Biochem Biophys Res Commun. 1986 Jul 16;138(1):304-11. doi: 10.1016/0006-291x(86)90280-9.

Abstract

18,54-SF cells, which secrete rat insulin-like growth factor II (rIGF-II), have abundant type 2 IGF receptors. We have purified the type 2 receptor from these cells by solubilization of crude membranes in Triton X-100, followed by chromatography on agarose-immobilized rIGF-II. A partially purified receptor preparation, obtained by chromatography of solubilized membranes over wheat germ agglutinin, was used to immunize a rabbit. The antibody generated both immunoprecipitates the type 2 receptor, and specifically inhibits IGF-II binding to a variety of rat tissues, including 18,54-SF cells, BRL-3A cells and placenta. The presence of abundant type 2 receptors on an rIGF-II-secreting cell line is consistent with an autocrine role for IGF-II in select cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies / immunology*
  • Cell Line
  • Chromatography, Affinity
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Insulin-Like Growth Factor I / metabolism
  • Kinetics
  • Lectins / metabolism
  • Octoxynol
  • Polyethylene Glycols
  • Receptors, Cell Surface / immunology
  • Receptors, Cell Surface / isolation & purification*
  • Receptors, Somatomedin
  • Solubility
  • Wheat Germ Agglutinins

Substances

  • Antibodies
  • Lectins
  • Receptors, Cell Surface
  • Receptors, Somatomedin
  • Wheat Germ Agglutinins
  • Polyethylene Glycols
  • Insulin-Like Growth Factor I
  • Octoxynol