Identification and intracellular location of MAGE-3 gene product

T. Kocher, E. Schultz-Thater, F. Gudat, C. Schaefer, G. Casorati, A. Juretic, T. Willimann, F. Harder, M. Heberer, G. C. Spagnoli

Research output: Contribution to journalArticle

Abstract

The human MAGE-3 gene encodes a melanoma antigenic epitope recognized by specific cytotoxic T lymphocytes, but its gene product has not been identified thus far. We produced a recombinant MAGE-3 gene product by expression cloning of the entire reading frame in the context of a fusion protein characterized by a 10-histidine tail, allowing purification by metal chelation on a nickel Sepharose column. The semipurified product was used to generate MAGE-3-specific monoclonal antibodies. One reagent could identify by immunoblotting the native MAGE-3 gene product as a M(r) 48,000 protein in lysates of cell lines showing evidence of MAGE-3 gene expression. No apparent cross-reactivity with recombinant or native MAGE-1 gene product was observed. Immunohistochemistry shows that, closely resembling the MAGE-1 gene product, MAGE-3 is a cytoplasmic protein.

Original languageEnglish
Pages (from-to)2236-2239
Number of pages4
JournalCancer Research
Volume55
Issue number11
Publication statusPublished - 1995

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

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    Kocher, T., Schultz-Thater, E., Gudat, F., Schaefer, C., Casorati, G., Juretic, A., Willimann, T., Harder, F., Heberer, M., & Spagnoli, G. C. (1995). Identification and intracellular location of MAGE-3 gene product. Cancer Research, 55(11), 2236-2239.