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E-Cadherin Antibody Staining Protocol for Immunohistochemistry
Description: Cadherins are a multifunctional family of Ca2+ dependent, transmembrane glycoproteins, which promote and maintain cell adhesion. They play an important role in cell-cell interaction, histogenesis and cellular transformation in virtually all multicellular organisms. The loss of E-Cadherin can result in the disruption of cell clustering. It is postulated in this matter that E-Cadherin may function as a tumor suppressor protein. In addition, the tissue specificity of cadherin subtypes are becoming valuable markers for identification and differential diagnosis of certain cancers (see Schofield, 1997 and Soler, 1997). The loss of E-Cadherin has been associated with metastasis and poor prognosis in invasive breast cancer, and can help differentiate between ductal and lobular neoplasms of the breast. E-cadherin has also been shown to be an independent predictor in the disease progression in bladder cancer.
Primary Antibody
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Name: Mouse Anti-Human E-Cadherin Antibody |
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Clone: 36B5 |
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Supplier: Millipore |
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Catalog Number: IHCR2123-6 |
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Dilution: Ready to Use. |
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Incubation Time/Temp: 60 min/room temperature |
Antigen Retrieval
| Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
| Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) |
| Heat/Cool Temperature: 95-100 ºC/room temperature |
| Heat/Cool Time: 20 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods |
Chromogen Substrate
| Reagent: DAB |
| Incubation Time/Temperature: 1-3 minutes/room temperature |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membrane |
| Images: Search image |
Additional Information:
| Species Reactivity: Human |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Normal breast |
| Negative Control: Omit primary antibody, isotype control, absorption control. |
| Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary. |
References: