Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Human PBMC (human primary peripheral blood mononuclear cell) cells labelling CD3D+CD3G+CD3 epsilon with ab318146 at 1/1000 (0.495 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in subsets of human PBMCs (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Anti-human CD4 mouse monoclonal antibody (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized MOLT-4 (human lymphoblastic leukemia T lymphoblast) cells labelling CD3D+CD3G+CD3 epsilon with ab318146 at 1/1000 (0.495 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in MOLT-4 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : Raji.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling CD3D+CD3G+CD3 epsilon with ab318146 at 1/5000 dilution (0.01ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti human CD19 conjugated to PE/Cy7. Fixed with 4% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab318146

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD3D+CD3G+CD3 epsilon with ab318146 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on T cell in human spleen. The section was incubated with ab318146 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IP

Supplier Data

Immunoprecipitation - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

CD3D+CD3G+CD3 epsilon was immunoprecipitated from 0.35 mg MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate with ab318146 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318146 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate
Lane 2 : ab318146 IP in MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab318146 in MOLT-4 whole cell lysate

All lanes:

Immunoprecipitation - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (ab318146) at 1/30 dilution

All lanes:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate with NFDM/TBST

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 6s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Mouse PBMC (mouse primary peripheral blood mononuclear cell) cells labelling CD3D+CD3G+CD3 epsilon with ab318146 at 1/1000 (0.495 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in subsets of mouse PBMCs (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Flow cytometric analysis of 4% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse PBMC cells labelling CD3D+CD3G+CD3 epsilon with ab318146 at 1/5000 dilution (0.01ug)/Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti mouse CD19 conjugated to PE/Cy7. Fixed with 4% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab318146.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling CD3D+CD3G+CD3 epsilon with ab318146 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on T cell in rat spleen. The section was incubated with ab318146 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized EL4 (mouse lymphoma T lymphocyte) cells labelling CD3D+CD3G+CD3 epsilon with ab318146 at 1/1000 (0.495 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in EL4 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : C2C12.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD3D+CD3G+CD3 epsilon with ab318146 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on T cell in mouse spleen. The section was incubated with ab318146 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IP

Supplier Data

Immunoprecipitation - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

CD3D+CD3G+CD3 epsilon was immunoprecipitated from 0.35 mg EL4 (mouse lymphoma T lymphocyte) whole cell lysate with ab318146 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab318146 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : EL4 (mouse lymphoma T lymphocyte) whole cell lysate
Lane 2 : ab318146 IP in EL4 (mouse lymphoma T lymphocyte) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab318146 in EL4 whole cell lysate

All lanes:

Immunoprecipitation - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (ab318146) at 1/30 dilution

All lanes:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate with NFDM/TBST

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Negative control : U-937, Raji, C2C12.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1-4 : 180 seconds Lane 5-7 : 103 seconds

All lanes:

Western blot - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (ab318146) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

MOLT-4 (human lymphoblastic leukemia T lymphoblast) whole cell lysate at 20 µg with NFDM/TBST

Lane 3:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg with NFDM/TBST

Lane 4:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg with NFDM/TBST

Lane 5:

EL4.IL-2 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg with NFDM/TBST

Lane 6:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg with NFDM/TBST

Lane 7:

C2C12 (mouse myoblast) whole cell lysate at 20 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 18-23 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Negative control : skeletal muscle.

The identity of the higher MW bands at approximately 75kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (ab318146) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 40 µg with NFDM/TBST

Lane 2:

Human skeletal muscle tissue lysate at 40 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 18-23 kDa,36 kDa

false

Exposure time: 81s

• WB

Supplier Data

Western blot - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Negative control : skeletal muscle and brain.

The identity of the higher MW bands at approximately 75kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (ab318146) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 40 µg with NFDM/TBST

Lane 2:

Mouse thymus tissue lysate at 40 µg with NFDM/TBST

Lane 3:

Mouse skeletal muscle tissue lysate at 40 µg with NFDM/TBST

Lane 4:

Mouse brain tissue lysate at 40 µg with NFDM/TBST

Lane 5:

Rat spleen tissue lysate at 40 µg with NFDM/TBST

Lane 6:

Rat thymus tissue lysate at 40 µg with NFDM/TBST

Lane 7:

Rat brain tissue lysate at 40 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 18-23 kDa,23 kDa,36 kDa

false

Exposure time: 59s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CD3D+CD3G+CD3 epsilon with ab318146 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse cerebrum. The section was incubated with ab318146 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling CD3D+CD3G+CD3 epsilon with ab318146 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human cerebrum. The section was incubated with ab318146 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CD3D+CD3G+CD3 epsilon with ab318146 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on rat cerebrum. The section was incubated with ab318146 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• WB

Supplier Data

Western blot - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (AB318146)

This antibody cross-reacts with both human and mouse CD3D+CD3G+CD3 epsilon.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-CD3D+CD3G+CD3 epsilon antibody [RM1128] (ab318146) at 1/1000 dilution

Lane 1:

HEK-293T cells transfected with an empty vector containing a His tag, whole cell lysate at 20 µg with NFDM/TBST

Lane 2:

HEK-293T cells transfected with a human CD3E vector containing a His tag, whole cell lysate at 4 µg with NFDM/TBST

Lane 3:

HEK-293T cells transfected with a human CD3G vector containing a His tag, whole cell lysate at 4 µg with NFDM/TBST

Lane 4:

HEK-293T cells transfected with a human CD3D vector containing a His tag, whole cell lysate at 4 µg with NFDM/TBST

Lane 5:

HEK-293T cells transfected with a mouse CD3E vector containing a His tag, whole cell lysate at 20 µg with NFDM/TBST

Lane 6:

HEK-293T cells transfected with a mouse CD3D vector containing a His tag, whole cell lysate at 20 µg with NFDM/TBST

Lane 7:

HEK-293T cells transfected with a mouse CD3G vector containing a His tag, whole cell lysate at 20 µg with NFDM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 18-25 kDa,36 kDa

false

Exposure time: 6s