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

Name: c-erbB-2 Oncoprotein Antibody

Clone: Polyclonal, Rabbit anti-Human

Supplier: Dako

Catalog Number: A0485

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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