CD31 Antibody Staining Protocol for Immunohistochemistry


Description: CD31 is a transmembrane glycoprotein, 130-140 kDa, also know as platelet-endothelium cell adhesion molecule (PECAM-1). CD31 is ligand for CD38 and plays a role in thrombosis and angiogenesis. CD31 is strongly expressed in endothelial cells and weakly expressed in megakaryocytes, platelets, occasional plasma cells, lymphocytes (espc. marginal zone B-cells, peripheral T-cells) and neutrophils. JC/70A reacts with a glycoprotein of about 100 kDa in endothelial cells and 130 kDa in platelets and can be used for the immunohistochemical detection of normal and tumor tissues.


Primary Antibody

Name: CD31 Antibody

Clone: JC/70A, Mouse anti-Human

Supplier: Dako

Catalog Number: M0823

Dilution: 1:20 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: For best results, incubate primary antibody for 2 hours/room temperature or overnight at 4 C.

Antigen Retrieval (Proteinase K digestion does not work for this antibody)
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 Chromogen Substrate
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Membrane/cytoplasmic of endothelial cells
Images: Search image

Additional Information:
Species Reactivity: Human
Fixation: Formalin fixed paraffin sections, acetone fixed frozen sections
Positive Control: Human 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


1. Not need to perform antigen retrieval on acetone fixed frozen sections

2. This antibody does not have cross reactivity with mouse and rat