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

Name: Vimentin Antibody

Clone: V9, Mouse monoclonal

Supplier: Invitrogen

Catalog Number: 18-0052

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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