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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
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Name: CD31 Antibody |
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Clone: JC/70A, Mouse anti-Human |
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Supplier: Dako |
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Catalog Number: M0823 |
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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. |
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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: