Characterization of a murine monoclonal antibody that detects a C-terminal fragment of the raf oncogene product

S. L. Giardina, S. M. Storm, D. L. Longo, B. J. Mathieson, U. Rapp, L. Varesio

Research output: Contribution to journalArticle

Abstract

A murine mAb, STEGI 1, was generated against a 30-kDa raf protein purified from an Escherichia coli expression vector. Immunoblot analysis confirmed that this antibody recognized the original immunizing protein as well as a 44- to 48-kDa protein from several raf-transported cell lines. Immunoprecipitation experiments isolated a 48-kDa protein from a cell line transfected with a c-raf construct as well as from normal NIH 3T3 fibroblasts. Parallel experiments with polyvalent antiserum prepared against E. coli-derived v-raf (C terminus)-precipitated proteins with apparent M(r) of 48 and 74 kDa, as had been described previously. Immunofluorescence flow cytometry of raf-transformed cell lines revealed intense intracytoplasmic staining. Thist staining was specifically inhibited by preincubation of STEGI 1 with purified raf 30-kDa protein. It should now be possible to more easily assess the role of the raf oncogene product in malignant transformation.

Original languageEnglish
Pages (from-to)3528-3533
Number of pages6
JournalJournal of Immunology
Volume140
Issue number10
Publication statusPublished - 1988

    Fingerprint

ASJC Scopus subject areas

  • Immunology

Cite this

Giardina, S. L., Storm, S. M., Longo, D. L., Mathieson, B. J., Rapp, U., & Varesio, L. (1988). Characterization of a murine monoclonal antibody that detects a C-terminal fragment of the raf oncogene product. Journal of Immunology, 140(10), 3528-3533.