Integrin alpha3 beta1 Antibody Staining Protocol for Immunohistochemistry

 

Description: Integrins are a family of dimeric proteins that mediate cell-cell and extracellular matrix adhesion. Each VLA subtype (VLA-1, VLA-2, VLA-3, VLA-4, VLA-5) is composed of a distinct alpha subunit noncovalently associated with a common beta subunit, as an alpha-beta dimmer. Integrin VLA-3 (very late antigen, alpha-3/beta-1) is a cell surface receptor that binds to fibronectin, laminin, collagen type I and VI. VLA-3 is typically restricted in normal tissue but is widely expressed in many cultured tumor cells and correlates with the degree of invasiveness of malignant melanoma cells. This antibody works well with acetone-fixed, frozen sections.

 

Primary Antibody

Name: Integrin alpha3 beta1 Antibody

Clone: M-K1D2, Mouse anti-Human

Supplier: Chemicon

Catalog Number: MAB1992

Dilution: 1:500 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: Antigen retrieval is not needed

Buffer/pH value: N/A

Heat/Cool Temperature: N/A

Heat/Cool Time: N/A

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: Basement membrane

Images: Search image

Additional Information:

Species Reactivity: Human

Fixation: Acetone-fixed, frozen sections. Not suitable for formalin fixed, paraffin embedded tissue sections.

Positive Control: Skin, renal carcinoma

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

 

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