c-Met (Hepatocyte Growth Factor Receptor) Antibody Staining Protocol for Immunohistochemistry


Description: The c-MET gene encodes a transmembrane tyrosine kinase identified as the receptor for a polypeptide known as hepatocyte growth factor (HGF). HGF has been shown to exert a pleiotropic activity on several cell types mainly of epithelial origin. It is a powerful mitogen for hepatocytes and also stimulates the growth of other cell types including kidney tubular cells, keratinocytes and endothelial cells. Other cell types known to express c-MET include hepatocytes, microglial cells in white matter and astrocytes.


Primary Antibody

Name: c-Met Antibody

Clone: 8F11, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-cMET

Dilution: 1:20 - 1:30 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:
Tissue Type: Prostate epithelium
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections.
Positive Control: Prostate epithelium
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


There is no need to perform antigen retrieval for frozen sections.