Detection of mouse tyrosinase with a monoclonal antibody MAT-1 against human tyrosinase.
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.
Suzuki
S
S
POLA R&D Laboratories, Yokohama, Japan.
Takimoto
H
H
Masui
S
S
Kato
T
T
Shibata
K
K
Tomita
Y
Y
Shibahara
S
S
Nakano
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D015153:Blotting, Western; D005591:Chemical Fractionation; D006801:Humans; D007527:Isoenzymes / Q000276:immunology; D008545:Melanoma / Q000201:enzymology / Q000276:immunology / Q000473:pathology; D051379:Mice; D008970:Molecular Weight; D014442:Monophenol Monooxygenase / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D014442:Monophenol Monooxygenase