Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

This data was developed using ab237707, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon cancer labelling CD208 with ab281573 at 1/1200 (B), CD3E with ab237707 at 1/1200 dilution (C) and CD19 with ab237772 at 1/1200 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-CD208 (red; Opal™570), anti-CD3E (green; Opal™520) and anti-CD19 (magenta; Opal™690) on human colon cancer.

Panel B : anti-CD208 staining dendritic cells in human colon cancer.

Panel C : anti-CD3E staining T lymphocytes in human colon cancer.

Panel D : anti-CD19 staining B lymphocytes of human colon cancer.

The section was incubated in three rounds of staining : in the order ab281573, ab237707, and ab237772 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

Supplier Data

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

This data was developed using ab237707, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil labelling CD208 with ab281573 at 1/1200 (B), CD3E with ab237707 at 1/1200 dilution (C) and CD19 with ab237772 at 1/1200 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-CD208 (red; Opal™570), anti-CD3E (green; Opal™520) and anti-CD19 (magenta; Opal™690) on human tonsil.
Panel B : anti-CD208 staining dendritic cells in human tonsil.
Panel C : anti-CD3E staining T lymphocytes in human tonsil.
Panel D : anti-CD19 staining B lymphocytes of human tonsil.

The section was incubated in three rounds of staining : in the order ab281573, ab237707, and ab237772 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

Supplier Data

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

This data was developed using ab237707, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon labelling CD208 with ab281573 at 1/1200 (B), CD3E with ab237707 at 1/1200 dilution (C) and CD19 with ab237772 at 1/1200 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-CD208 (red; Opal™570), anti-CD3E (green; Opal™520) and anti-CD19 (magenta; Opal™690) on human colon.

Panel B : anti-CD208 staining dendritic cells in human colon.

Panel C : anti-CD3E staining T lymphocytes in human colon.

Panel D : anti-CD19 staining B lymphocytes of human colon.

The section was incubated in three rounds of staining : in the order ab281573, ab237707, and ab237772 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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

Immunohistochemical analysis of paraffin-embedded human gastric carcinoma tissue labeling CD3 epsilon with ab237707 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the infiltrating T lymphocytes in the human gastric carcinoma is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab237707 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween 20 permeabilized human PBMCs (peripheral blood mononuclear cells) labeling CD3 epsilon with ab237707 at 1/500 (Right compared with a Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Left). Goat Anti-Rabbit IgG Fc (Alexa Fluor^®488) preadsorbed (ab150097), at 1/5000 dilution was used as the secondary antibody.

Cells were surface stained with anti-CD3 epsilon conjugated to Alexa Fluor^® 647. Then fixed with 2% PFA for 10min followed by intracellular staining rabbit IgG (Left) or ab237707 (Right).

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

Formalin-fixed, paraffin-embedded human tonsil tissue stained for CD3 epsilon using ab237707 at 0.5 μg/mL in immunohistochemcial analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

Formalin-fixed, paraffin-embedded human NSCLC tissue stained for CD3 epsilon using ab237707 at 0.5 μg/mL in immunohistochemcial analysis.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling CD3 epsilon with ab237707 at 1/55 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in Jurkat cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

PBS only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD3 epsilon with ab237707 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the human tonsil is observed. Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

The section was incubated with ab237707 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

This data was developed using ab237707, the same antibody clone in a different buffer formulation.

Fluorescence multiplex immunohistochemical analysis of human tonsil (formalin-fixed paraffin-embedded section). Merged staining of Panel A : merged staining of anti-TCR beta (green; Opal™520) and anti-CD3E (red; Opal™570) on human tonsil, Panel B : anti-TCR beta staining T lymphocytes of human tonsil, Panel C : anti-CD3E staining T lymphocytes of human tonsil. DAPI was used as a counter stain. Followed by Opal Polymer HRP Ms + Rb. 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. The section was incubated in three rounds of staining : in the order of ab315324, ab237707 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

This data was developed using ab237707, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver cancer labelling CD208 with ab281573 at 1/1200 (B), CD3E with ab237707 at 1/1200 dilution (C) and CD19 with ab237772 at 1/1200 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-CD208 (red; Opal™570), anti-CD3E (green; Opal™520) and anti-CD19 (magenta; Opal™690) on human liver cancer.

Panel B : anti-CD208 staining dendritic cells in human liver cancer.

Panel C : anti-CD3E staining T lymphocytes in human liver cancer.

Panel D : anti-CD19 staining B lymphocytes of human liver cancer.

The section was incubated in three rounds of staining : in the order ab281573, ab237707, and ab237772 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.

• IP

Supplier Data

Immunoprecipitation - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

CD3 epsilon was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab237707 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237707 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1 : Jurkat whole cell lysate 10 μg (Input).
Lane 2 : ab237707 IP in Jurkat whole cell lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab237707 in Jurkat whole cell lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

All lanes:

Immunoprecipitation - Anti-CD3 epsilon antibody [CAL54] (<a href='/en-us/products/primary-antibodies/cd3-epsilon-antibody-cal54-ab237707'>ab237707</a>)

Predicted band size: 23 kDa

Observed band size: 21 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-CD3 epsilon antibody [CAL54] - BSA and Azide free (AB251594)

CD3 epsilon was immunoprecipitated from 0.35 mg of human thymus lysate with ab237707 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237707 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

Lane 1 : Human thymus lysate 10 μg (Input).
Lane 2 : ab237707 IP in human thymus lysate.
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab237707 in human thymus lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.
Exposure time : 30 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237707).

All lanes:

Immunoprecipitation - Anti-CD3 epsilon antibody [CAL54] (<a href='/en-us/products/primary-antibodies/cd3-epsilon-antibody-cal54-ab237707'>ab237707</a>)

Predicted band size: 23 kDa

Observed band size: 21 kDa

false