Cytokeratin HMW/34βE12

Cytokeratin 34βE12 is a High Molecular Weight cytokeratin that reacts with all squamous and ductal epithelium and stains carcinomas. This antibody recognizes cytokeratins 1, 5, 10, and 14 that are found in complex epithelia. Cytokeratin 34βE12 shows no reactivity with hepatocytes, pancreatic acinar cells, proximal renal tubules or endometrial glands; there has been no reactivity with cells derived from simple epithelia. Nerve cells, glial cells and mesenchymal tissue such as blood vessels containing only non-keratin types of intermediate filaments are not labelled; however, reactivity with smooth-muscle cells has been occasionally observed.

Mesenchymal Tumors, Lymphomas, Melanomas, Neural Tumors and Neuroendocrine Tumors are unreactive with this antibody. Cytokeratin 34βE12 has been shown to be useful in distinguishing Prostatic Adenocarcinoma from Hyperplasia of the Prostate.

Cytokeratin 34βE12 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.