 Methods & Techniques For Histologists and Immunohistochemists |
c-erbB-2/HER-2/neu Antibody Staining Protocol for Immunohistochemistry
Description: c-erbB-2 is a receptor tyrosine kinase of the c-erbB family. It is closely related instructure to the epidermal growth factor receptor. c-erbB-2 oncoprotein is detectable in a proportion of breast and other adenocarconomas, as well as transitional cell carcinomas. In the case of breast cancer, expression determined by immunohistochemistry has been shown to be associated with poor prognosis.
Primary Antibody
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Name: c-erbB-2/HER-2/neu Antibody |
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Clone: SP3, Rabbit monoclonal |
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Supplier: Lab Vision |
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Catalog Number: RM-9103-S |
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Dilution: 1:100 |
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Incubation Time/Temp: 60 minutes/room temperature |
Antigen Retrieval
| Device: Steamer |
| Buffer/pH value: Tris-EDTA/pH 9.0 |
| 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: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membrane |
| Images: Search image |
Additional Information:
| Tissue Type: Breast carcinoma |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Breast 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 |
References: