Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using the same antibody clone in a different buffer formulation ( ab237721).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse lymph node tissue staining Neutrophil Elastase with ab310335 at a 1/500 ( 1.076 µg/ml) dilution, CD19 with ab245235 at a 1/1000 ( 0.444 µg/ml) dilution and CD3 with ab237721 at a 1/2000 ( 0.264 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B : anti-Neutrophil Elastase staining neutrophils in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab310335, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse colon tissue staining Glycam1 with ab324912 at a 1/4000 ( 0.126 μg/ml) dilution, ab237721 anti-CD3 used at 1/2000 dilution ( 0.26 μg/ml) and ab315346 anti-CD37 used at 1/2000 dilution (0.257 μg/ml).

Panel A : anti-Glycam1 (green; Opal™520), anti-CD3 (magenta; Opal™690), anti-CD37 (gray; Opal™570) on mouse colon.

Panel B : anti-Glycam1 staining HEV (highly endothelial venules) in mouse colon.

Panel C : anti-CD3 staining T lymphocytes in mouse colon.

Panel D : anti-CD37 staining B lymphocytes in mouse colon.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324912, ab237721 and ab315346 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse lymph node staining of NCR1 with ab233558 at a 1/500 (1.062 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-NCR1 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse lymph node.
Panel B : anti-NCR1 staining natural killer cells in mouse lymph node.
Panel C : anti-CD19 staining B lymphocytes in mouse lymph node..
Panel D : anti-CD3 staining T lymphocytes in mouse lymph node.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab233558, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lymph node tissue staining Glycam1 with ab324912 at a 1/4000 ( 0.126 μg/ml) dilution, ab237721 anti-CD3 used at 1/2000 dilution ( 0.26 μg/ml) and ab315346 anti-CD37 used at 1/2000 dilution (0.257 μg/ml).

Panel A : anti-Glycam1 (green; Opal™520), anti-CD3 (magenta; Opal™690), anti-CD37 (gray; Opal™570) on mouse lymph node.

Panel B : anti-Glycam1 staining HEV (highly endothelial venules) in mouse lymph node.

Panel C : anti-CD3 staining T lymphocytes in mouse lymph node.

Panel D : anti-CD37 staining B lymphocytes in mouse lymph node.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab324912, ab237721 and ab315346 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Immunohistochemistry analysis of (Formalin/PFA-fixed paraffin-embedded sections) Rat lymph node tissue labelling MAdCAM1 with ab309487 at 1 : 100 dilution (B), CD3 epsilon with ab237721at 1 : 2000 dilution (C) and CD20 with ab64088 at 1 : 100 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-MAdCAM (green; Opal™520), anti-CD3 (gray; Opal™690) and anti-CD20 (magenta; Opal™570) on rat lymph node.

Panel B : anti-MAdCAM staining high endothelial venules in rat lymph node.

Panel C : ant-CD3 staining B lymphocytes in rat lymph node.

Panel D : ant-CD20 staining B lymphocytes in rat lymph node.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab309487, ab237721 and ab64088 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse small intestine tissue staining CD68 with ab283654 at a 1/100 dilution, ab245235 anti-CD19 used at 1/1000 dilution and ab237721 anti-CD3 used at a 1/2000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD68 (magenta; Opal^™520), anti-CD19 (green; Opal^™690) and anti-CD3 (grey; Opal^™570) on mouse small intestine.
Panel B : anti-CD68 staining macrophages in mouse small intestine.
Panel C : anti-CD19 staining B lymphocytes in mouse small intestine.
Panel D : anti-CD3 staining T lymphocytes in mouse small intestine.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab283654, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^®RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse colon tissue staining CD68 with ab283654 at a 1/100 dilution, ab245235 anti-CD19 used at 1/1000 dilution and ab237721 anti-CD3 used at a 1/2000 dilution. Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-CD68 (magenta; Opal^™520), anti-CD19 (green; Opal^™690) and anti-CD3 (grey; Opal^™570) on mouse colon.
Panel B : anti-CD68 staining macrophages in mouse colon.
Panel C : anti-CD19 staining B lymphocytes in mouse colon.
Panel D : anti-CD3 staining T lymphocytes in mouse colon.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab283654, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^®RX instrument with an Opal^™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Immunohistochemistry analysis of (Formalin/PFA-fixed paraffin-embedded sections) Mouse lymph node tissue labelling MAdCAM1 with ab309487 at 1 : 100 dilution (B), CD3 epsilon with ab237721at 1 : 2000 dilution (C) and CD20 with ab64088 at 1 : 100 dilution (D). Opal Polymer HRP Ms + Rb was used as a secondary antibody, and DAPI was used for a nuclear counter stain. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Panel A : merged staining of anti-MAdCAM (green; Opal™520), anti-CD3 (gray; Opal™690) and anti-CD20 (magenta; Opal™570) on mouse lymph node.

Panel B : anti-MAdCAM staining high endothelial venules in mouse lymph node.

Panel C : ant-CD3 staining B lymphocytes in mouse lymph node.

Panel D : ant-CD20 staining B lymphocytes in mouse lymph node.

Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab309487, ab237721 and ab64088 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using the same antibody clone in a different buffer formulation (ab237721).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse lymph node tissue staining CD23 with ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with ab237721 at 1/8000 dilution (0.07 µg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD23 (green; Opal™520), anti-CD19 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on mouse lymph node.
Panel B : anti-CD23 staining B lymphocytes in mouse lymph node.
Panel C : anti-CD19 staining B lymphocytes in mouse lymph node.
Panel D : anti-CD3 staining T lymphocytes in mouse lymph node.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315289, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using the same antibody clone in a different buffer formulation (ab237721).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse thymus tissue staining CD23 with ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with ab237721 at 1/8000 dilution (0.07 µg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD23 (green; Opal™520), anti-CD19 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on mouse thymus.
Panel B : anti-CD23 staining B lymphocytes in mouse thymus.
Panel C : anti-CD19 staining B lymphocytes in mouse thymus.
Panel D : anti-CD3 staining T lymphocytes in mouse thymus.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315289, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen staining of NCR1 with ab233558 at a 1/500 (1.062 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-NCR1 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse spleen.
Panel B : Anti-NCR1 staining natural killer cells in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab233558, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using the same antibody clone in a different buffer formulation ( ab237721).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse bone marrow tissue staining Neutrophil Elastase with ab310335 at a 1/500 ( 1.076 µg/ml) dilution, CD19 with ab245235 at a 1/1000 ( 0.444 µg/ml) dilution and CD3 with ab237721 at a 1/2000 ( 0.264 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B : anti-Neutrophil Elastase staining neutrophils in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab310335, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using the same antibody clone in a different buffer formulation (ab237721).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded mouse spleen tissue staining CD23 with ab315289 at a 1/2000 dilution (0.253 µg/ml), CD19 with ab245235 at 1/1000 dilution (0.444 µg/ml), and CD3 with ab237721 at 1/8000 dilution (0.07 µg/ml), followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-CD23 (green; Opal™520), anti-CD19 (magenta; Opal™570) and anti-CD3 (yellow; Opal™690) on mouse spleen.
Panel B : anti-CD23 staining B lymphocytes in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab315289, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse bone marrow staining of F4/80 with ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse bone marrow.
Panel B : anti-F4/80 staining macrophages in mouse bone marrow.
Panel C : anti-CD19 staining B lymphocytes in mouse bone marrow.
Panel D : anti-CD3 staining T lymphocytes in mouse bone marrow.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300421, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse thymus staining of F4/80 with ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse thymus.
Panel B : anti-F4/80 staining macrophages in mouse thymus.
Panel C : anti-CD19 staining B lymphocytes in mouse thymus.
Panel D : anti-CD3 staining T lymphocytes in mouse thymus.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300421, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse thymus staining of NCR1 with ab233558 at a 1/500 (1.062 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-NCR1 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse thymus.
Panel B : anti-NCR1 staining natural killer cells in mouse thymus.
Panel C : anti-CD19 staining B lymphocytes in mouse thymus.
Panel D : anti-CD3 staining T lymphocytes in mouse thymus.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab233558, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse lymph node staining of F4/80 with ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse lymph node.
Panel B : anti-F4/80 staining macrophages in mouse lymph node.
Panel C : anti-CD19 staining B lymphocytes in mouse lymph node.
Panel D : anti-CD3 staining T lymphocytes in mouse lymph node.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300421, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using the same antibody clone in a different buffer formulation ( ab237721).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse thymus tissue staining Neutrophil Elastase with ab310335 at a 1/500 ( 1.076 µg/ml) dilution, CD19 with ab245235 at a 1/1000 ( 0.444 µg/ml) dilution and CD3 with ab237721 at a 1/2000 ( 0.264 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B : anti-Neutrophil Elastase staining neutrophils in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab310335, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using the same antibody clone in a different buffer formulation ( ab237721).

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen tissue staining Neutrophil Elastase with ab310335 at a 1/500 ( 1.076 µg/ml) dilution, CD19 with ab245235 at a 1/1000 ( 0.444 µg/ml) dilution and CD3 with ab237721 at a 1/2000 ( 0.264 µg/ml) dilution followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-Neutrophil Elastase (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™690) on mouse spleen.Panel B : anti-Neutrophil Elastase staining neutrophils in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab310335, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

This data was developed using ab237721, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Mouse spleen staining of F4/80 with ab300421 at a 1/5000 (0.1 µg/ml) dilution, CD19 with ab245235 at 1/1000 (0.444 µg/ml) dilution and CD3 with ab237721 at 1/2000 ( 0.264 µg/ml) dilution.

Panel A : merged staining of anti-F4/80 (magenta; Opal™690), anti-CD19 (green; Opal™520) and anti-CD3 (gray; Opal™570) on mouse spleen.
Panel B : anti-F4/80 staining macrophages in mouse spleen.
Panel C : anti-CD19 staining B lymphocytes in mouse spleen.
Panel D : anti-CD3 staining T lymphocytes in mouse spleen.
Nuclear DNA was labelled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300421, ab245235 and ab237721 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0 epitope retrieval solution2) for 20 mins.

• IP

Lab

Immunoprecipitation - Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (AB251607)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).

All lanes:

Immunoprecipitation - Anti-CD3 epsilon antibody [CAL57] (<a href='/en-us/products/primary-antibodies/cd3-epsilon-antibody-cal57-ab237721'>ab237721</a>)

Predicted band size: 23 kDa

false