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

Name: c-erbB-2 Oncoprotein Antibody

Clone: CBE1, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CBE1

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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