Detection of mouse tyrosinase with a monoclonal antibody MAT-1 against human tyrosinase

Satoshi Suzuki, Hiroyuki Takimoto, Shigeki Masui, Tomomi Kato, Koushi Shibata, Yasushi Tomita, Shigeki Shibahara, Hiroyuki Nakano

Research output: Contribution to journalArticlepeer-review

Abstract

In this study we explored the possible application of MAT-1, which has been established as a monoclonal antibody against human tyrosinase, for detection of mouse tyrosinase. The MAT-1 reacted with B16 mouse melanoma cells, but not with tyrosinase-negative NIH-3T3 mouse fibroblasts. In western blot analysis of the large granule fraction (LGF) of B16 cells, MAT-1 detected a single protein of 80 kDa, whose size was close to that of human tyrosinase detected with MAT-1 in extracts of human melanocytes. Furthermore, the 80 kDa band that was detected with MAT-1 in the LGF of B16 cells was also detected by DOPA reaction. In order to confirm that the protein detected with MAT-1 is tyrosinase, a transient expression assay was carried out. When mouse tyrosinase or mouse tyrosinase-related protein 1, which shares high homology with human tyrosinase, was transiently expressed in tyrosinase-negative K1735 mouse melanoma cells by cDNA transfection, MAT-1 reacted only with the cells expressing mouse tyrosinase. These results indicate that MAT-1 specifically reacts with mouse tyrosinase.

Original languageEnglish
Pages (from-to)298-303
Number of pages6
JournalPigment Cell Research
Volume9
Issue number6
DOIs
Publication statusPublished - 1996 Dec

Keywords

  • Melanoma cell
  • Transient expression assay
  • Western blot analysis

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science
  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

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