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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
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Name: CD31 Antibody |
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Clone: 1A10, Mouse anti-Human |
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Supplier: Zymed |
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Catalog Number: 18-2425 |
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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. |
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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 |
Counterstain
| Reagent: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| 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 |
Notes:
Not need to perform antigen retrieval on acetone fixed frozen sections |
References: