CD31 Antibody Staining Protocol for Immunohistochemistry


Description: This antibody reacts with rat Pecam-1. Antibody gives positive reaction at high dilutions against recombinant rat PECAM-1. This antibody also cross reacts with Pig endothelial cells and platelets as determined by histochemistry on acetone fixed frozen sections and flow cytometry. Reactivity with other species not determined. Non reactive with human CD31.


Primary Antibody

Name: CD31 Antibody

Clone: TLD-3A12, Mouse anti-Rat

Supplier: Research Diagnostics

Catalog Number: RDI-RTCD31-3A

Dilution: 1:50 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: 60min/room temperature

Antigen Retrieval
Device: Incubator
Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101)
Heat/Cool Temperature: 37 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
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic of endothelial cells and platelets
Images: Search image

Additional Information:
Tissue Type: Tonsil
Fixation: Zinc fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Tonsil, cardiac tissues, skin, kidney
Negative Control: Omit primary antibody, isotype control or 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 for acetone fixed frozen sections