ALK-1

Anaplastic Lymphoma Kinase (ALK) was originally discovered as a NPM (Nucleophosmin)-ALK fusion protein. The ALK gene is on chromosome 2. Upon translocation between chromosome 2 and chromosome 5 t(2;5), the ALK gene fuses with the NPM gene. The chimeric product (NPM ALK) resulting from t(2;5) translocation is a protein of 80 kDa with the N terminal portion of NPM linked to the complete intracellular portion of ALK.

This antibody recognizes a human p80 protein, identified as a hybrid of the Anaplastic Lymphoma Kinase (ALK) gene and the Nucleophosmin (NPM) gene resulting from the t(2;5)(p23;q35) translocation found in a third of Large-Cell Lymphomas. ALK-1 is detected in 60% of Anaplastic Large-Cell Lymphomas and has proven to indicate a better prognosis in the ALK-1 (+) group.

Anti-ALK-1 is a rabbit 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. Cataldo KA, et al. Am J Surg Pathol. 1999:32(1):1386-1392.
2. Nakamura S, Shiota M, et al. Am J Surg Pathol. 1997:21(12):1420-1432.
3. Falini B, Bigerna B, et al. Am J Pathol. 1998: 153(3)Sept. 875-886.

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.