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Vimentin Antibody Staining Protocol for Immunohistochemistry
Description: Vimentin is an intermediate filament that is ubiquitously expressed in mesenchymal cells such as fibroblasts, smooth muscle cells, and endothelium. Vimentin is useful as part of an antibody panel for differential diagnostic purposes, such as differentiating soft tissue tumors. Positive tissue: Endometrial carcinoma or malignant fibrous histiocytoma.
Specificity: Mouse Anti-Vimentin (Clone: V9) is specific for vimentin, and does not recognize any other cellular or intermediate filament proteins such as desmin or glial fibrillary acidic proteins.
Primary Antibody
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Name: Vimentin Antibody |
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Clone: V9, Mouse monoclonal |
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Supplier: Invitrogen |
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Catalog Number: 18-0052 |
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Dilution: 1:1000 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: Tonsil, skin, smooth muscle |
| 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 |
References: