E-Cadherin Antibody Staining Protocol for Immunohistochemistry


Description: This monoclonal antibody reacts with human epithelial cadherin (E-cadherin, uvomorulin, Cell-CAM 120/80), a 120 kDa transmembrane glycoprotein. Reduction and/or loss of E-cadherin expression in carcinomas correlates positively with tumor cell invasion and metastasis. Altered expression of E-cadherin is common in the progression of cancers including breast carcinoma, lung carcinoma, gastric tumors, hepatocellular carcinoma, and prostatic tumors. It has recently been reported that E-cadherin is a useful marker for differentiating lobular lesions (negative) from ductal invasive lesions (positive) in breast cancer.


Primary Antibody

Name: E-Cadherin Antibody

Clone: 4A2C7, Mouse anti-Human

Supplier: Zymed

Catalog Number: 18-0223

Dilution: 1:100 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 minutes/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 ºC - 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: Mainly cellular membrane, some cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Non-neural epithelia
Fixation: Formalin fixed paraffin sections
Positive Control: Non-neural epithelia
Negative Control: Omit primary antibody, isotype control or 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


Not tested on frozen sections



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2.   Parker C, Rampaul RS, Pinder SE, Bell JA, Wencyk PM, Blamey RW, Nicholson RI, Robertson JF (2001) E-cadherin as a prognostic indicator in primary breast cancer. Br J Cancer. 85(12):1958-63. PubMed Abstract

3.   Rubin MA, Mucci NR, Figurski J, Fecko A, Pienta KJ, Day ML (2001) E-cadherin expression in prostate cancer: a broad survey using high-density tissue microarray technology. Hum Pathol. 32(7):690-7. PubMed Abstract

4.   Alami J, B R Williams and H Yeger (2003) Differential expression of E-cadherin and ß catenin in primary and metastatic Wilms’s tumours. Molecular Pathology. 56:218-225