Myogenin

Myogenin is a transcription factor active in muscles. In particular, it is a myogenic regulatory factor. Myogenin is a member of a family of myogenic regulatory genes, which includes MyoD, myf5 and MRF4. These genes encode a set of transcription factors which are essential for muscle development. Expression of myogenin is restricted to cells of skeletal-muscle origin. It is, therefore, a useful marker for tumors of the muscle lineage, being strongly expressed in Alveolar Rhabdomyosarcomas.

Anti-myogenin labels the nuclei of myoblasts in developing muscle tissue, and is expressed in tumor cell nuclei of Rhabdomyosarcoma and some Leiomyosarcomas. Positive nuclear staining may occur in Wilm’s Tumor.

Myogenin 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.

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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.