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c-erbB-2 Oncoprotein Antibody Staining Protocol for Immunohistochemistry
Description: The c-erbB-2 oncoprotein is closely-related in structure to the epidermal growth factor receptor and is a member of a large family of cell surface growth factor receptors. c-erbB-2 oncoprotein is reported to be detectable in a proportion of breast and other adenocarcinomas as well as transitional cell carcinomas. c-erbB-2 oncoprotein is present in a wide variety of cell types in a range of normal human fetal and adult tissues, including breast, stomach and ovary.
Primary Antibody
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Name: c-erbB-2 Oncoprotein Antibody |
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Clone: CBE1, Mouse anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-CBE1 |
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Dilution: 1:20 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: Overnight/4 ºC |
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:
| Tissue Type: Breast, stomach, ovary |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Cultured SK-BR-3 cells |
| 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: