CD3 Antibody Staining Protocol for Immunohistochemistry

 

Description: The alpha-beta heterodimeric T-cell antigen receptor binds antigen in association with major histocompatibility complex proteins on host cell surfaces. These 2 disulfide-linked glycoproteins, TCRA and TCRB are associated on T-cell surfaces with a complex of proteins called CD3 (formerly T3). Human CD3 consists of at least 4 proteins: gamma, delta , epsilon, and zeta . The CD3-gamma/CD3-delta gene pair is within 300 kb of the CD3-epsilon gene and therefore these genes form a tightly linked cluster on 11q23. The clustering may be significant in terms of their simultaneous activations during T-cell development.

 

Primary Antibody

Name: CD3 IHC Antibody

Clone: Mouse Monoclonal: CA-3

Supplier: IHC World

Catalog Number: IW-MA1015

Dilution: Ready to Use

Incubation Time/Temp: 60 min/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:

Species Reactivity: Human, rat

Fixation: Formalin fixed paraffin embedded sections or acetone fixed frozen sections.

Positive Control: Human tonsil

Negative Control: Omit primary antibody, isotype control, absorption control

Blocking: Normal serum blocking is not needed for this RTU antibody; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary

 

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