CD31 Antibody Staining Protocol for Immunohistochemistry


Description: CD31 is useful as a marker for tumor angiogenesis as it stains blood vessels. It is also a more sensitive marker than CD34, UEA-1 and von Willebrand factor for malignant vascular endothelium. This monoclonal antibody (clone 1A10) reacts with CD31. Immunogen source: Recombinant fusion protein representing a portion of the extracellular domain of CD31. The CD31 clone 1A10 is optimized for use on formalin fixed paraffin embedded tissues.


Primary Antibody

Name: CD31 Antibody

Clone: 1A10, Mouse anti-Human

Supplier: Zymed

Catalog Number: 18-2425

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 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: 30 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: Membrane of endothelial cells
Images: Search image

Additional Information:
Tissue Type: Tonsil
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Tonsil, skin, liver, kidney
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


Not need to perform antigen retrieval on acetone fixed frozen sections