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CD3 (T Cell) Antibody Staining Protocol for Immunohistochemistry
Description: The CD3 molecule consists of five different polypeptide chains with molecular weights ranging from 16 to 28 kD. The five chains are designated gamma, delta, epsilon, zeta and eta. This antibody is specific for the non-glycosylated epsilon chain of the CD3 molecule. The CD3 complex is involved in signal transduction to the T cell interior following antigen recognition. The CD3 antigen is first detectable in early thymocytes and its appearance probably represents one of the earliest signs of commitment to the T cell lineage.
Primary Antibody
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Name: CD3, T Cell Antibody |
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Clone: Rabbit anti-Human |
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Supplier: Dako |
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Catalog Number: A0452 |
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Dilution: 1:50 - 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: 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: Membrane of T cells |
| Images: Search image |
Additional Information:
| Species Reactivity: Human, cynomolgus macaque, mouse |
| Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
| Positive Control: Tonsil, spleen, thymus |
| 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 for acetone fixed frozen sections |
References: