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Epithelial Membrane Antigen (EMA) Antibody Staining Protocol for Immunohistochemistry
Description: Epithelial membrane antigen (EMA), is a mucin-like glycoprotein. Antibody to EMA has been shown useful as a pan- epithelial marker for detecting early metastatic loci of carcinoma in the bone marrow or liver.
Primary Antibody
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Name: EMA Ab-3 Antibody |
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Clone: E29, Mouse anti-Human |
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Supplier: Lab Vision |
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Catalog Number: MS-741 |
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Dilution: 1:50 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: No antigen retrieval is needed. |
| Buffer/pH value: N/A |
| Heat/Cool Temperature: N/A |
| Heat/Cool Time: N/A |
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: Cytoplasmic and cell membrane |
| Images: Search image |
Additional Information:
| Tissue Type: Breast cancer, Colon carcinoma |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Breast cancer, Colon carcinoma |
| 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: