CD2 Antibody Staining Protocol for Immunohistochemistry

Description: CD2 is a 50 kDa single-chain type I transmembrane glycoprotein, originally defined as the sheep erythrocyte receptor. CD2 primarily binds CD58 on antigen-presenting cells and induces costimulatory signals in T cells. CD2 is one of the earliest T-cell lineage-restricted antigens to appear during T-cell differentiation, and it is expressed on most thymocytes, peripheral T cells and NK cells. In addition, CD2 has been shown to be expressed on approximately 50% of thymic B cells, while the expression on mature B cells is controversial.

Primary Antibody

Name: CD2 Antibody

Clone: MT910, Mouse anti-Human

Supplier: Dako

Catalog Number: M0720

Dilution: 1:100 using IHC-Tek^TM 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-Tek^TM Epitope Retrieval Steamer Set (Cat# IW-1102)

Buffer/pH value: IHC-Tek^TM Epitope Retrieval Solution (Cat# IW-1100)

Heat/Cool Temperature: 95-100 ºC/room temperature

Heat/Cool Time: 45 minutes/20 minutes

Detection Methods

Standard Method: ABC Method or LSAB Method

Enhanced Method: Polymeric Methods is preferred for this antibody

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, or acetone fixed frozen 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

Notes:

There is no need to perform antigen retrieval for acetone fixed frozen sections

References: