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CD16 Antibody Staining Protocol for Immunohistochemistry
Description: CD16 is a low affinity Fc receptor of complexed IgG, Fc/gamma RIII and expressed on NK cells, granulocytoes, activated macrophages and a subset of T cells. CD16 exists both as a glycosyl-phosphatidylinositol-anchored protein in polymorphonuclear cells and as a transmembrane protein in NK cells. This antigen will help detect CD16 and NK cells in a variety of tissues.
Primary Antibody
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Name: CD16 Antibody |
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Clone: 2H7, Mouse anti-Human |
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Supplier: Vector Labs |
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Catalog Number: VP-C333 |
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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: 60min/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: 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: Gill's Hematoxylin or Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Membrane staining of macrophage, neutrophils, NK cells and granulocytes |
| Images: Search image |
Additional Information:
| Tissue Type: Tonsil |
| Fixation: Formalin fixed paraffin sections |
| Positive Control: Tonsil |
| 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 effective on fixed frozen sections |
References: