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CD3 zeta 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 for zeta chain.
Primary Antibody
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Name: CD3 zeta Antibody |
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Clone: Rabbit anti-Human |
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Supplier: GeneTex |
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Catalog Number: GTX17061 |
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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: 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: Cell Membrane |
| Images: Search image |
Additional Information:
| Species Activity: Human, mouse, rat |
| 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 |
References: