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CD163 Antibody Staining Protocol for Immunohistochemistry
Description: CD163 is cell surface glycoprotein, a 175kD molecule also known as ED2. CD163 is expressed by approximately 50% of peritoneal macrophages, a subset of splenic macrophages, and by macrophages in most other tissues. However, it is not expressed by monocytes, alveolar macrophages or microglial cells.
Primary Antibody
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Name: CD163 Antibody |
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Clone: ED2, Mouse anti-Rat |
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Supplier: AbD Serotec |
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Catalog Number: MCA342R |
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Dilution: 1:500 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: 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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
| Species Reactivity: Rat |
| Tissue Type: Kupffer cells of liver, dendrocytes of spleen |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Kupffer cells of liver, dendrocytes of spleen |
| 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 |
Notes:
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Not need to perform antigen retrieval on acetone fixed frozen sections |
References: