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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
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Name: E-Cadherin Antibody |
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Clone: 4A2C7, Mouse anti-Human |
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Supplier: Zymed |
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Catalog Number: 18-0223 |
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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. |
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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 |
Counterstain
| Reagent: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
Notes:
Not tested on frozen sections |
References:
1. Shnayder Y, Kuriakose MA, Yee H, Chen FA, DeLacure MD, Xue XN, Jagirdar J (2001) Adhesion molecules as prognostic factors in nasopharyngeal carcinoma. Laryngoscope. 111(10):1842-6. PubMed Abstract
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