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

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
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

Notes:

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

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

 

References: