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

Name: CD3, T Cell Antibody

Clone: Rabbit anti-Human

Supplier: Dako

Catalog Number: A0452

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.

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

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

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


Not need to perform antigen retrieval for acetone fixed frozen sections