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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
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Name: c-Met Antibody |
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Clone: 8F11, Mouse anti-Human |
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Supplier: Novocastra Labs |
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Catalog Number: NCL-cMET |
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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. |
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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: 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 |
Notes:
There is no need to perform antigen retrieval for frozen sections. |
References: