Integrin alpha2 beta1 Antibody Staining Protocol for Immunohistochemistry

 

Description: This antibody reacts with the collagen receptor VLA-2 (α 2 β 1) integrin complex and it is suitable for immunohistochemistry in frozen tissue sections. It does not detect α2 integrin subunit by Western blot presumably due to conformational properties of the epitope involved. The epitope for BHA2.1 is dependent on the presence of the (I) domain of α 2 integrin subunit and does not bind VLA-2 variant which lacks the I-domain. BHA2.1 inhibits VLA-2 mediated cell adhesion to collagen and laminin; complete inhibition can generally be achieved at a final concentration of 10 ug/ml.

 

Primary Antibody

Name: Integrin alpha2 beta1 (collagen receptor) Antibody

Clone: BHA2.1, Mouse anti-Human

Supplier: Chemicon

Catalog Number: MAB1998, MAB1998Z

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

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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