Anti-TBX3 antibody [EPR28578-38]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TBX3 antibody [EPR28578-38] (AB322199)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet (B) TBX3 knockout HAP1 cell pellet tissue labeling TBX3 with ab322199 at 1/2000 (0.253 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on A : wild-type HAP1 cell pellet, no staining on B : TBX3 knockout HAP1 cell pellet. The section was incubated with ab322199 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-TBX3 antibody [EPR28578-38] (AB322199)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling TBX3 with ab322199 at 1/1000 (0.506 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast carcinoma (PMID : 30697731). The section was incubated with ab322199 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-TBX3 antibody [EPR28578-38] (AB322199)

Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling TBX3 with ab322199 at 1/1000 (0.506 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human endometrium. The section was incubated with ab322199 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-TBX3 antibody [EPR28578-38] (AB322199)

Immunohistochemical analysis of paraffin-embedded Human placenta tissue labeling TBX3 with ab322199 at 1/1000 (0.506 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human placenta. The section was incubated with ab322199 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-TBX3 antibody [EPR28578-38] (AB322199)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling TBX3 with ab322199 at 1/1000 (0.506 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on human pancreas (PMID : 27632063). The section was incubated with ab322199 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-TBX3 antibody [EPR28578-38] (AB322199)

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

The identity of the bands lower than 75 kDa are unknown.

Exposure time : Lane 1 : 180 seconds; Lane 2 : 103 seconds.

All lanes:

Western blot - Anti-TBX3 antibody [EPR28578-38] (ab322199) at 1/1000 dilution

Lane 1:

Human adrenal gland tissue lysate at 20 µg

Lane 2:

Human placenta tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 79 kDa

false

• WB

Supplier Data

Western blot - Anti-TBX3 antibody [EPR28578-38] (AB322199)

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

The identity of the lower MW band at approximately 60 kDa is unknown.

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.

All lanes:

Western blot - Anti-TBX3 antibody [EPR28578-38] (ab322199) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

TBX3 knockout HAP1 whole cell lysate at 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 1/20000 dilution

Observed band size: 79 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-TBX3 antibody [EPR28578-38] (AB322199)

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

The identity of the lower MW band at approximately 60 kDa is unknown.

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.

All lanes:

Western blot - Anti-TBX3 antibody [EPR28578-38] (ab322199) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeting TBX3 whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 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 1/20000 dilution

Observed band size: 79 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-TBX3 antibody [EPR28578-38] (AB322199)

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

Low expression : HL-60.

The identity of the bands lower than 75 kDa are unknown.

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

All lanes:

Western blot - Anti-TBX3 antibody [EPR28578-38] (ab322199) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 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 1/20000 dilution

Observed band size: 79 kDa,36 kDa

false

Exposure time: 70s

• Dot

Supplier Data

Dot Blot - Anti-TBX3 antibody [EPR28578-38] (AB322199)

Dot blot analysis of TBX3 using ab322199 at 1 : 1000 (0.506 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : 293T cells transfected with a human TBX3 expression vector containing a myc-His-tag®, whole cell lysate
Lane2 : 293T cells transfected with a human TBX2 expression vector containing a myc-His-tag®, whole cell lysate

Exposure time : 180 seconds.

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

This antibody does not cross-react with overexpressed human TBX2 by dot blot.

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

All lanes:

Dot Blot - Anti-TBX3 antibody [EPR28578-38] (ab322199) at 1/1000 dilution

Lane 1:

293T cells transfected with a human TBX3 expression vector containing a myc-His-tag®, whole cell lysate

Lane 2:

293T cells transfected with a human TBX2 expression vector containing a myc-His-tag®, whole cell lysate

Secondary

All lanes:

Dot 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

false

Exposure time: 180s