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Related Experiment Videos

A sensitive tyrosinase method for human skin.

A Wittbjer1, G Odh, E Rosengren

  • 1Department of Pharmacology, University Hospital, Lund, Sweden.

Acta Dermato-Venereologica
|January 1, 1991
PubMed
Summary
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A new radioimmunoassay accurately measures tyrosinase enzyme levels in biological tissues. This method quantifies tyrosinase in skin samples, even with interfering substances present.

Area of Science:

  • Biochemistry
  • Immunology
  • Analytical Chemistry

Background:

  • Tyrosinase is a key enzyme in melanin production.
  • Accurate quantification of tyrosinase is crucial for understanding pigmentation disorders and melanoma.
  • Existing methods may be affected by inhibitors or oxidation in biological samples.

Purpose of the Study:

  • To develop a sensitive and quantitative assay for tyrosinase in biological tissues.
  • To establish a radioimmunoassay (RIA) method for tyrosinase analysis.
  • To validate the RIA for use with crude biological samples like skin.

Main Methods:

  • A competitive radioimmunoassay was developed using 125I-labelled human tyrosinase and antityrosinase antibodies.
  • Solubilized tyrosinase from melanoma and skin samples was used.

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  • The assay involved competition for antibody binding sites between labelled and unlabelled tyrosinase.

    • Main Results:

    • The radioimmunoassay demonstrated a sensitivity range of approximately 5-500 pmol/l.
    • The method was successfully applied to solubilized tyrosinase from 0.5 cm2 skin samples.
    • The RIA provided reliable tyrosinase quantification in crude samples, unaffected by inhibitors or oxidation.

    Conclusions:

    • A sensitive and specific competitive radioimmunoassay for tyrosinase has been successfully developed.
    • This RIA method is suitable for quantifying tyrosinase in biological tissues, including skin.
    • The assay offers an advantage for analyzing tyrosinase in complex samples where enzyme activity may be compromised.