Anti-TOX antibody [EPR28108-10]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOX antibody [EPR28108-10] (AB322259)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling TOX with ab322259 at 1/500 (1.004 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human skeletal muscle. The section was incubated with ab322259 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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOX antibody [EPR28108-10] (AB322259)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Daudi (human Burkitt's lymphoma lymphoblast, Left) / Raji (human Burkitt's lymphoma B lymphocyte, Right) cells labelling TOX with ab322259 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 was used as the secondary antibody.

Negative control : Daudi.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOX antibody [EPR28108-10] (AB322259)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling TOX with ab322259 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 was used as the secondary antibody.

Cells were stained with anti-CD8a conjugated to Pacific Blue. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOX antibody [EPR28108-10] (AB322259)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling TOX with ab322259 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 was used as the secondary antibody.

Cells were stained with anti-CD19 conjugated to PE/Cy7. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-TOX antibody [EPR28108-10] (AB322259)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling TOX with ab322259 at 1/50 dilution.

Confocal image showing cytoplasmic staining in Raji cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).

Negative control : Daudi.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Anti-alpha Tubulin mouse monoclonal antibody (ab7291) was used to counterstain tubulin at 1/1000 dilution (Magenta).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOX antibody [EPR28108-10] (AB322259)

Immunohistochemical analysis of paraffin-embedded (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a TOX expression vector containing a Myc-His tag, (B) HEK-293T transfected with a TOX2 expression vector containing a Myc-His tag, (C) HEK-293T transfected with a TOX3 expression vector containing a Myc-His tag, (D) HEK-293T transfected with a TOX4 expression vector containing a Myc-His tag and (E) HEK-293T transfected with empty vector containing a Myc-His tag labeling TOX with ab322259 at 1/2000 (0.251 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Positive staining on (A) HEK-293T (human epithelial cell line from embryonic kidney) transfected with a TOX expression vector containing a Myc-His tag, no staining on (B) HEK-293T transfected with a TOX2 expression vector containing a Myc-His tag, (C) HEK-293T transfected with a TOX3 expression vector containing a Myc-His tag, (D) HEK-293T transfected with a TOX4 expression vector containing a Myc-His tag and (E) HEK-293T transfected with empty vector containing a Myc-His tag.
The section was incubated with ab322259 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-TOX antibody [EPR28108-10] (AB322259)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling TOX with ab322259 at 1/500 (1.004 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human spleen (PMID : 32106280). The section was incubated with ab322259 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-TOX antibody [EPR28108-10] (AB322259)

Immunohistochemical analysis of paraffin-embedded Human thymus tissue labeling TOX with ab322259 at 1/500 (1.004 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human thymus (PMID : 32106280). The section was incubated with ab322259 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-TOX antibody [EPR28108-10] (AB322259)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling TOX with ab322259 at 1/500 (1.004 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human tonsil (PMID : 32106280). The section was incubated with ab322259 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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOX antibody [EPR28108-10] (AB322259)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized B16-F10 (mouse skin melanoma cell, Left) / EL4 (mouse lymphoma T lymphocyte, Right) cells labelling TOX with ab322259 at 1/5000 compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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

Low expression : B16-F10.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOX antibody [EPR28108-10] (AB322259)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse PBMCs (mouse primary peripheral blood mononuclear cells) cells labelling TOX with ab322259 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 was used as the secondary antibody.

Cells were stained with anti-CD8 conjugated to Brilliant Violet 421. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOX antibody [EPR28108-10] (AB322259)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling TOX with ab322259 at 1/500 (1.004 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse pancreas (PMID : 32106280). The section was incubated with ab322259 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-TOX antibody [EPR28108-10] (AB322259)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling TOX with ab322259 at 1/500 (1.004 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse skeletal muscle. The section was incubated with ab322259 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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOX antibody [EPR28108-10] (AB322259)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Rat PBMC (Rat peripheral blood mononuclear cell) cells labelling TOX with ab322259 at 1/50 compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

Cells were stained with anti-CD8a conjugated to Alexa Fluor®647. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOX antibody [EPR28108-10] (AB322259)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse PBMCs (mouse primary peripheral blood mononuclear cells) cells labelling TOX with ab322259 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control.

Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 was used as the secondary antibody.

Cells were stained with anti-CD14 conjugated to Brilliant Violet 421. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TOX antibody [EPR28108-10] (AB322259)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling TOX with ab322259 at 1/500 (1.004 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse kidney. The section was incubated with ab322259 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

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-TOX antibody [EPR28108-10] (AB322259)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Rat PBMC (Rat peripheral blood mononuclear cell) cells labelling TOX with ab322259 at 1/50 compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

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

Cells were stained with anti-CD11b/c conjugated to PE. Then fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our antibody.

• WB

Supplier Data

Western blot - Anti-TOX antibody [EPR28108-10] (AB322259)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, cells/tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

Exposure time : Lane 1 : 180 seconds; Lane 2 : 37 seconds; Lane 3 : 125 seconds.

All lanes:

Western blot - Anti-TOX antibody [EPR28108-10] (ab322259) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 57 kDa,63 kDa

false

• WB

Supplier Data

Western blot - Anti-TOX antibody [EPR28108-10] (AB322259)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : Daudi (PMID : 32106280), AsPC-1, A549.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

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

Exposure time : Lane 1 : 180 seconds; Lanes 2-5 : 15 seconds; Lane 6 : 48 seconds.

All lanes:

Western blot - Anti-TOX antibody [EPR28108-10] (ab322259) at 1/1000 dilution

Lane 1:

KARPAS-299 (human t cell lymphoma cell) whole cell lysate at 20 µg

Lanes 2 and 6:

NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

AsPC-1 (human Pancreas epithelial cell) whole cell lysate at 20 µg

Lane 4:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Daudi (human burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 57 kDa,63 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-TOX antibody [EPR28108-10] (AB322259)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : skeletal muscle.

To minimize protein degradation, tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

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

All lanes:

Western blot - Anti-TOX antibody [EPR28108-10] (ab322259) at 1/1000 dilution

Lane 1:

Mouse thymus tissue lysate at 20 µg

Lane 2:

Mouse skeletal muscle tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 57 kDa,63 kDa,36 kDa

false

Exposure time: 6s