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CD2 Antibody Staining Protocol for Immunohistochemistry
Description: CD2 is a 50 kDa single-chain type I transmembrane glycoprotein, originally defined as the sheep erythrocyte receptor. CD2 primarily binds CD58 on antigen-presenting cells and induces costimulatory signals in T cells. CD2 is one of the earliest T-cell lineage-restricted antigens to appear during T-cell differentiation, and it is expressed on most thymocytes, peripheral T cells and NK cells. In addition, CD2 has been shown to be expressed on approximately 50% of thymic B cells, while the expression on mature B cells is controversial.
Primary Antibody
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Name: CD2 Antibody |
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Clone: MT910, Mouse anti-Human |
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Supplier: Dako |
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Catalog Number: M0720 |
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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: 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: 45 minutes/20 minutes |
Detection Methods
| Standard Method: ABC Method or LSAB Method |
| Enhanced Method: Polymeric Methods is preferred for this antibody |
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 |
| Images: Search image |
Additional Information:
| Tissue Type: Tonsil |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen 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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There is no need to perform antigen retrieval for acetone fixed frozen sections |
References: