New EIA technique for tyrosinase in human melanocytes and its application

Nagaoki Wakisaka, Hajime Inoue, Takehiko Nagase, Kyo Ichi Matsuzaki, Masaki Aihara, Norio Kumagai

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)


The expression of tyrosinase in melanocytes relates to skin pigmentation or depigmentation. Although many types of drugs with whitening effects are well known, neither the definite effect nor the mechanism underlying the effect has been elucidated. In this study, we attempted to develop the rapid and simple EIA technique for tyrosinase protein, then this technique was applied to normal human cultured melanocytes. When primary antibody and tyrosinase were incubated in non-coated 96-well microtitre plates for 48 hours at 4 °C, then the solution in tyrosinase-coated plate was further incubated for another 1 hour at 37°C. Thus the best results were obtained. The developed EIA system could detect authentic tyrosinase until 0.1-1.0 ng/mL. This EIA technique could also be applied to human cultured melanocytes. The melanocytes cultured with endothelin-1 induced tyrosinase like immune reactive protein. The protein induction with endothelin-1 was suppressed by BQ 123, ETa receptor antagonists. The simple EIA technique developed for tyrosinase may give a clue to determination of the onset mechanisms underlying pigmental diseases of the skin as well as the mechanisms underlying the effects of various whitening drugs.

Original languageEnglish
Pages (from-to)PL-1-PL-6
JournalLife Sciences
Issue number1
Publication statusPublished - 1999 Nov 26
Externally publishedYes


  • BQ123
  • EIA
  • Endothelin-1
  • Melanocytes
  • Tyrosinase

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)


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