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Vimentin Antibody Staining Protocol for Immunohistochemistry
Description: Vimentin is a 55 kDa intermediate filament (IF) protein, which form part of the cytoskeleton of vertebrate cells. Among the five classes of IFs vimentin belongs to class III and is expressed in the cytoplasma of cells of mesenchymal origin. Being the predominant IF protein in mesenchymal cells vimentin was initially believed to be a useful marker for distinguishing non-epithelial from epithelial derived nepolasms. The co-expression of intermediate filaments, particularly vimentin and cytokeratin, has now been demonstrated in a variety of normal cells/tissues and in neoplastic lesions, necessitating the use of a panel of antibodies in differential tumour diagnosis.
Primary Antibody
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Name: Mouse Anti-Human Vimentin Antibody |
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Clone: Vim 3B4 |
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Supplier: DakoCytomation |
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Catalog Number: M7020 |
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Dilution: 1:800 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 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: Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Human |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Normal breast |
| 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. if IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) is used, there is no serum blocking step needed. |
References: