Tryptase

Tryptase is the most abundant secretory granule-derived serine proteinase contained in mast cells and has recently been used as a marker for mast cell activation. It is involved in allergenic response and is suspected to act as a mitogen for fibroblast lines. Elevated levels of serum tryptase occur in both anaphylactic and anaphylactoid reactions, but a negative test does not exclude anaphylaxis. Mast cells contain a number of preformed chemical mediators such as histamine, chymase, carboxypeptidase and proteolytic tryptase.

Human mast cell tryptase is considered to be an important marker of mast cell activation as well as an important mediator of inflammation. Anti-tryptase is a good marker for mast cells, basophils, and their derivatives.

Tryptase 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. Aoki, et al. Int Arch Allergy Immunol. 2003;130(3):216-223
2. Ghott, et al. Am J Surg Pathol. 2003;27(7):1013-1019
3. Fiorucci L, et al. Cell Mol Life Sci. 2004;61(11):1278-1295

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.