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c-erbB-2 Oncoprotein Antibody Staining Protocol for Immunohistochemistry
Description: Polyclonal Rabbit Anti-Human c-erbB-2 Oncoprotein is intended for use in immunocytochemistry. The antibody labels normal epithelial cells, which, however, generally express c-erbB-2 protein at a very low level. It is a useful tool for the identification of overexpression of cerbB-2 oncoprotein in a variety of epithelial neoplasms, for example subsets of breast carcinomas, pulmonary adenocarcinomas, colorectal adenocarcinomas, pulmonary squamous and gastric adenocarcinomas, transitional cell carcinomas of the urinary bladder, and endometrial adenocarcinomas. Differential identification is aided by the results from a panel of antibodies. Interpretation of results must be made within the context of the patient’s clinical history and other diagnostic tests by a certified professional.
Primary Antibody
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Name: c-erbB-2 Oncoprotein Antibody |
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Clone: Polyclonal, Rabbit anti-Human |
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Supplier: Dako |
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Catalog Number: A0485 |
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Dilution: 1:500 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: 95-100 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: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membrane |
| Images: Search image |
Additional Information:
| Species Activity: Human |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Breast carcinomas. c-erbB-2 oncoprotein is a normal tissue component and the antibody may display a weak labelling of normal epithelial cells. Squamous epithelium in the esophagus and tonsil may in some cases show moderate staining. Prostatic gland tissue has also been found moderately positive. |
| 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: