Recombinant Anti-CD3 epsilon antibody [CAL57] (ab237721)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD3 epsilon with ab237721 at 1/2000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the T lymphocytes in the human tonsil. The section was incubated with ab237721 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CD3 epsilon with ab237721 at 1/2000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the T lymphocytes in the human colon. The section was incubated with ab237721 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse thymus tissue labeling CD3 epsilon with ab237721 at 1/500 dilution (green), followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Positive staining on T cell surface (PMID: 8490660) is observed. Counterstained with DAPI (blue). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit used at a 1/1,000 dilution. Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).

The expression pattern is consistent with what described in the literature. (PMID: 19542373).

Blocking/Dilution buffer: 5% NFDM/TBST.

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized mouse splenocytes labeling CD3 epsilon with ab237721 at 1/500 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. 

Cells were surface stained with anti-CD3 conjugated to Alexa Fluor^® 647. Then fixed with 2% PFA for 10min and intracellularly stained with rabbit IgG (Left) or ab237721 (Right). They recognize the same cell population. 

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized human peripheral blood mononuclear cell (PBMC) labeling CD3 epsilon with ab237721 at 1/500 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150097), at 1/5000 dilution was used as the secondary antibody. 

Cells were surface stained with anti-CD3 conjugated to Alexa Fluor^® 647. Then fixed with 2% PFA for 10min and intracellularly stained with rabbit IgG (Left) or ab237721 (Right). They recognize the same cell population. 

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD3 epsilon with ab237721 at 1/2000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the T lymphocytes in the rat spleen. The section was incubated with ab237721 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3 epsilon with ab237721 at 1/2000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the T lymphocytes in the mouse spleen. The section was incubated with ab237721 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

CD3 epsilon was immunoprecipitated from 0.35 mg EL4.IL2 (Mouse lymphoma T lymphocyte) whole cell lysate with ab237721 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237721 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.
Lane 1: EL4.IL2 whole cell lysate 10 µg (Input).
Lane 2: ab237721 IP in EL4.IL2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237721 in EL4.IL2 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 5 seconds.

Formalin-fixed, paraffin-embedded mouse spleen tissue stained for CD3 epsilon using ab237721 at 0.5 µg/ml in immunohistochemical analysis.