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

Name: c-erbB-2/HER-2/neu Antibody

Clone: SP3, Rabbit monoclonal

Supplier: Lab Vision

Catalog Number: RM-9103-S

Dilution: 1:100 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

Counterstain
Reagent: 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

 

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