CD3 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. This antibody recognizes T cells in thymus, bone marrow, peripheral lymphoid tissue and blood and detect both normal and neoplastic T cells.

 

Primary Antibody

Name: CD3 Antibody

Clone: PS1, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CD3-PS1

Dilution: 1:200 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

Counterstain
Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Results:
Staining Pattern: Membrane
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

 

References: