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

Name: Mouse Anti-Human E-Cadherin Antibody

Clone: 36B5

Supplier: Millipore

Catalog Number: IHCR2123-6

Dilution: Ready to Use.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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.