Anti-T-bet / Tbx21 antibody [EPR27094-16]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin's tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on human non-Hodgkin's lymphoma (T cell). The section was incubated with ab307193 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 Solution 2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : No staining on human cerebrum. The section was incubated with ab307193 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 Solution 2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on human tonsil. The section was incubated with ab307193 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 Solution 2) for 20 mins

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) cells labelling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Live cells were stained with anti-CD56 conjugated to BV421. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab307193.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) cells labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : CD19+ population. Live cells were stained with anti-CD19 conjugated to Alexa Fluor® 647. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab307193.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer (NK-92®) cell with no GFP tag) cells labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2µg/ml dilution (Green). Confocal image showing nuclear staining in No-GFP-CD16.NK-92 cell line. Negative control : K562. 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 2.5µg/ml dilution (Red). 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 2µg/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K562 (human chronic myelogenous leukemia lymphoblast, Left) / No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag, Right) cells labelling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : K562.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse peripheral blood mononuclear cell (PBMC) cells labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : CD19+ population. Live cells were stained with anti-CD19 conjugated to Alexa Fluor® 647. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab307193.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Mouse splenocytes cells labeling T-bet / Tbx21 with ab307193 at 1/500 dilution (0.1µg)/ Right (Red) compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Live cells were stained with anti-CD3 conjugated to Alexa Fluor® 647. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or ab307193.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on mouse spleen. The section was incubated with ab307193 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 Solution 2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Immunohistochemical analysis of paraffin-embedded Mouse Burkitt's lymphoma tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on mouse Burkitt's lymphoma. The section was incubated with ab307193 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 Solution 2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling T-bet / Tbx21 with ab307193 at 1/500 (1.044 µg/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on rat spleen. The section was incubated with ab307193 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 Solution 2) for 20 mins

• WB

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Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : K-562 In lane 1, the lysate was freshly made and used for Western Blotting immediately to minimize protein degradation. The identity of the bands below 50 kDa are unknown. Exposure time : 15 seconds

All lanes:

Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution

Lane 1:

No-GFP-CD16.NK-92 (human malignant non-Hodgkins lymphoma natural killer cell) whole cell lysate 20 μg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate 20 μg

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: 60 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The lysate was freshly made and used for Western Blotting immediately to minimize protein degradation. The identity of the bands below 50 kDa are unknown. Exposure time : 103 seconds

All lanes:

Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution

All lanes:

Rat spleen tissue lysate 20 μg

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: 60 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (AB307193)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The lysate was freshly made and used for Western Blotting immediately to minimize protein degradation. Exposure time : 180 seconds

All lanes:

Western blot - Anti-T-bet / Tbx21 antibody [EPR27094-16] (ab307193) at 1/1000 dilution

All lanes:

Mouse spleen tissue lysate 20 μg

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: 60 kDa

false

Exposure time: 180s