Fibronectin Antibody Staining Protocol for Immunohistochemistry


Description: Fibronectin is a glycoprotein that has functions associated with cell adhesion and migration. It is found primarily in basement membranes and is a key extracellular matrix component. This antibody will help detect fibronectin in normal and uncharacterized tissues.


Primary Antibody

Name: Fibronectin Antibody

Clone: 568, Mouse anti-Human

Supplier: Vector Labs

Catalog Number: VP-F705

Dilution: 1:100 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 minutes/room temperature

Antigen Retrieval
Device: Incubator
Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101)
Heat/Cool Temperature: 37 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: Extracellular connective tissue matrix, basement membranes
Images: Search image

Additional Information:
Tissue Type: Skin, kidney
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections, or cell smears.
Positive Control: Normal kidey (extracellular labeling of connective tissue matrix), basement membrane of skin, blood vessels, smooth muscles, ganglia and muscle fibers.
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


No need to perform antigen retrieval on frozen sections