Vimentin Antibody Staining Protocol for Immunohistochemistry


Description: This monoclonal antibody reacts with the intermediate filament protein vimentin present in cells of mesenchymal origin. Broad interspecies cross-reactivity has been observed with human, rat, and chicken vimentin. No reaction with other closely related intermediate filament proteins including desmin, keratin, neurofilament, and glial fibrillary acid protein was observed in immunoblotting analysis. Vim 3B4 labels cells of mesenchymal origin and their tumors. Reactive cells include normal and neoplastic lymphoid cells, endothelial cells, smooth and striated muscle cells, as well as Schwann cells. Other intermediate filament proteins may be coexpressed, such as desmin in vascular smooth muscle cells and glial fibrillary acidic protein in some glial cells.


Primary Antibody

Name: Vimentin Antibody

Clone: VIM 3B4, Mouse monoclonal

Supplier: Dako

Catalog Number: M7020

Dilution: 1:400 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, rat, chicken
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen 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