Epidermal Growth Factor Receptor (EGFr) Antibody Staining Protocol for Immunohistochemistry

 

Description: This monoclonal antibody reacts with the extracellular domain of epidermal growth factor receptor (170 kDa). Clone 31G7 is useful in detecting the majority of squamous cell carcinomas that arise from both squamous epithelium and metaplastic squamous epithelium. Positive tissue: Breast carcinoma.

 

Primary Antibody

Name: Mouse Anti-EGFr Antibody

Clone: 31G7

Supplier: invitrogen

Catalog Number: 08-1205

Dilution: Ready to Use.

Incubation Time/Temp: 60 min/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/Cytoplasmic
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections
Positive Control: Breast carcinoma
Negative Control: Omit primary antibody, isotype control, 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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