E-Cadherin

Cadherins are a class of transmembrane proteins. They play an important role in cell adhesion by ensuring cells within tissues are bound together. E-Cadherin is an adhesion protein that is expressed in cells of epithelial lineage. It stains positively in glandular epithelium as well as Adenocarcinomas of the lung and G.I. tract, and ovary. E-Cadherin has been useful in distinguishing Adenocarcinoma from Mesothelioma. It has also been shown to be positive in some Thyroid Carcinomas. It can be used to differentiate Ductal Carcinomas (positive for E-Cadherin) from Lobular Breast Carcinomas.

Loss of E-Cadherin function or expression has been implicated in cancer progression and metastasis. E-Cadherin downregulation decreases the strength of cellular adhesion within a tissue, resulting in an increase in cellular motility. This may then allow cancer cells to cross the basement membrane and invade surrounding tissues. Loss of E-Cadherin expression has been suggested as a poor prognostic sign in Breast Carcinoma and Non-Small Cell Lung Carcinomas.

E-Cadherin 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.

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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.