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

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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

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Oncogene Proteins
Monoclonal Antibodies
Proteins
Staining and Labeling
Cell Line
Transformed Cell Line
Escherichia coli Proteins
Immunoprecipitation
Fluorescent Antibody Technique
Immune Sera
Flow Cytometry
Fibroblasts
Escherichia coli
Antibodies

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.

Characterization of a murine monoclonal antibody that detects a C-terminal fragment of the raf oncogene product. / Giardina, S. L.; Storm, S. M.; Longo, D. L.; Mathieson, B. J.; Rapp, U.; Varesio, L.

In: Journal of Immunology, Vol. 140, No. 10, 1988, p. 3528-3533.

Research output: Contribution to journalArticle

Giardina, SL, Storm, SM, Longo, DL, Mathieson, BJ, 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, vol. 140, no. 10, pp. 3528-3533.
Giardina, S. L. ; Storm, S. M. ; Longo, D. L. ; Mathieson, B. J. ; Rapp, U. ; Varesio, L. / Characterization of a murine monoclonal antibody that detects a C-terminal fragment of the raf oncogene product. In: Journal of Immunology. 1988 ; Vol. 140, No. 10. pp. 3528-3533.
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