Tyrosinase

Tyrosinase is an enzyme that catalyzes the oxidation of phenols (such as tyrosine) and is widespread in plants and animals. Tyrosinase is a copper-containing enzyme present in plant and animal tissues that catalyzes the production of melanin and other pigments from tyrosine by oxidation. The gene for Tyrosinase is regulated by the Microphthalmia-associated transcription factor. A mutation in the tyrosinase gene leads to impaired tyrosinase production resulting in Type I Oculocutaneous Albinism, a hereditary disease that affects 1 in 17,000 people in the U.S.

Anti-Tyrosinase has been found to be quite specific for melanotic lesions such as Malignant Melanoma and Melanotic Neurofibroma. Essentially no carcinomas express this marker.

Anti-Tyrosinase is a mouse monoclonal antibody derived from cell culture supernatant that is concentrated, dialyzed, filter sterilized and diluted in buffer pH 7.5, containing BSA and sodium azide as a preservative.

1. Kaufmann O, et al. Mod Pathol. 1998;Aug;11(8):740-6
2. Busam KJ, et al. Am J Dermatopathol. 2000;June;22(3):237-41
3. Jungbluth AA, et al. Pathol Res Pract. 2000;196(4):235-42
4. Meije CB, et al. J Pathol. 2000;Apr;190(5):572-8

1. Cut and mount 3-4 micron formalin-fixed paraffin-embedded tissues on positive charged slides.
2. Air dry for 2 hours at 58° C.
3. Deparaffinize, dehydrate and rehydrate tissues.
4. Subject tissues to heat epitope retrieval using a suitable retrieval solution such as ImmunoDNA Retriever with Citrate (BSB 0020-BSB 0023) or EDTA (BSB 0030-BSB 0033).
5. Any of three heating methods may be used:
   • Electric Pressure Cooker
     Place standoff rack at base of pressure cooker. Add 1-2 inches of distilled water to the pressure cooker and turn heat to high, and incubate for 15 minutes. Open and immediately transfer slides to room temperature.
   • Water Bath Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a water bath set at 95°-99° C. Incubate for 30-60 minutes.
   • Conventional Steamer Method
     Place tissues/slides in a pre-warmed staining dish or coplin jar containing the ImmunoDNA Retriever with Citrate or EDTA in a Steamer, cover and steam for 30-60 minutes.
6. After heat treatment, transfer slides in ImmunoDNA Retriever with Citrate or EDTA to room temperature and let stand for 15-20 minutes.
7. Wash slides with IHC wash buffer or DI water.
8. Continue IHC staining protocol.