Generation of a murine monoclonal antibody that detects the fos oncogene product

Steven L. Giardina, Stuart W. Evans, Lucia Gandino, Frank A. Robey, E. Bonvini, Dan L. Longo, Luigi Varesio

Research output: Contribution to journalArticle

Abstract

A hybridoma producing a monoclonal antibody (MoAB) recognizing both the cellular and viral forms of fos has been generated by somatic cell hybridization techniques from spleen cells of mice immunized with a synthetic peptide corresponding to amino acids 128-152, a consensus region, of both the v-fos and c-fos oncogene products. Three proteins with molecular weights of 55,000, 44,000, and 42,000 were detected by immunoblotting. While MoAB 2G9C3 failed to immunoprecipitate fos from Finkel-Biskis-Jenkins murine osteosarcoma-virus-in-fected fibroblasts, both the 55,000 v-fos protein and the 39,000 cellular protein were coprecipitated using polyvalent rabbit antibodies to the same peptide. Whereas no cell surface membrane expression of fos was detected, after membrane permeabilization by a brief exposure to lysolecithin it was possible to specifically detect internal fos by immunofluorescence flow cytometry. Immunohistochemical staining of FBJ virus-infected cells revealed intense, nuclear staining.

Original languageEnglish
Pages (from-to)109-116
Number of pages8
JournalAnalytical Biochemistry
Volume161
Issue number1
DOIs
Publication statusPublished - Feb 15 1987

Keywords

  • monoclonal antibodies
  • oncogene
  • synthetic peptide

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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  • Cite this

    Giardina, S. L., Evans, S. W., Gandino, L., Robey, F. A., Bonvini, E., Longo, D. L., & Varesio, L. (1987). Generation of a murine monoclonal antibody that detects the fos oncogene product. Analytical Biochemistry, 161(1), 109-116. https://doi.org/10.1016/0003-2697(87)90659-2