Anti-Brd4 antibody [EPR29186-23]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR29186-23] (AB322063)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Brd4 with ab322063 at 1/100 (5.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon. The section was incubated with ab322063 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-Brd4 antibody [EPR29186-23] (AB322063)

Immunohistochemical analysis of paraffin-embedded Human colon carcinoma tissue labeling Brd4 with ab322063 at 1/100 (5.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon carcinoma. The section was incubated with ab322063 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-Brd4 antibody [EPR29186-23] (AB322063)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Brd4 with ab322063 at 1/100 (5.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab322063 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-Brd4 antibody [EPR29186-23] (AB322063)

Immunohistochemical analysis of paraffin-embedded Rat kidney tissue labeling Brd4 with ab322063 at 1/100 (5.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat kidney. The section was incubated with ab322063 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-Brd4 antibody [EPR29186-23] (AB322063)

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Brd4 with ab322063 at 1/100 (5.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse kidney. The section was incubated with ab322063 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-Brd4 antibody [EPR29186-23] (AB322063)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling Brd4 with ab322063 at 1/100 (5.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse colon. The section was incubated with ab322063 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-Brd4 antibody [EPR29186-23] (AB322063)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Brd4 with ab322063 at 1/100 (5.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse testis. The section was incubated with ab322063 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-Brd4 antibody [EPR29186-23] (AB322063)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling Brd4 with ab322063 at 1/100 (5.11 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat testis. The section was incubated with ab322063 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-Brd4 antibody [EPR29186-23] (AB322063)

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

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-Brd4 antibody [EPR29186-23] (ab322063) 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 Brd4S 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 1/100000 dilution

Observed band size: 99 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Brd4 antibody [EPR29186-23] (AB322063)

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

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-Brd4 antibody [EPR29186-23] (ab322063) at 1/1000 dilution

All lanes:

K-562 (human chronic myelogenous leukemia lymphoblast) 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: 99 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Brd4 antibody [EPR29186-23] (AB322063)

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

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.

Exposure time : Lanes 1-2 : 26 seconds, Lanes : 3-4 : 92 seconds, Lane 5 : 37 seconds

All lanes:

Western blot - Anti-Brd4 antibody [EPR29186-23] (ab322063) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 5:

ES-D3 [D3] (mouse blastocyst-derived embryonic stem 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: 99 kDa

false

• WB

Supplier Data

Western blot - Anti-Brd4 antibody [EPR29186-23] (AB322063)

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

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.

All lanes:

Western blot - Anti-Brd4 antibody [EPR29186-23] (ab322063) at 1/1000 dilution

Lane 1:

Rat liver tissue lysate at 20 µg

Lane 2:

Rat testis tissue lysate at 20 µg

Lane 3:

Rat spleen tissue 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: 99 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Brd4 antibody [EPR29186-23] (AB322063)

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

In lanes 1-3, 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.

The identity of the lower MW band at approximately 37 kDa (in lane 1) is unknown.

All lanes:

Western blot - Anti-Brd4 antibody [EPR29186-23] (ab322063) at 1/1000 dilution

Lane 1:

Mouse liver tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Mouse placenta tissue 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: 99 kDa

false

Exposure time: 180s

• Dot

Supplier Data

Dot Blot - Anti-Brd4 antibody [EPR29186-23] (AB322063)

Dot blot analysis of Brd4 using ab322063 at 1 : 1000 (0.511 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Brd4S peptide a
Lane2 : Brd4S peptide b
Lane3 : BRD4L peptide
Lane4 : BAZ1A peptide
Lane5 : BRWD1 peptide
Lane6 : NMDE4 peptide
Lane7 : ZMYD8 peptide
Lane8 : B3A4 peptide

Exposure time : 180 seconds.

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

This antibody does not cross-react with human BRD4L, BAZ1A, BRWD1, NMDE4, ZMYD8 and B3A4.

All lanes:

Dot Blot - Anti-Brd4 antibody [EPR29186-23] (ab322063) at 1/1000 dilution

Lane 1:

Brd4S peptide a

Lane 2:

Brd4S peptide b

Lane 3:

BRD4L peptide

Lane 4:

BAZ1A peptide

Lane 5:

BRWD1 peptide

Lane 6:

NMDE4 peptide

Lane 7:

ZMYD8 peptide

Lane 8:

B3A4 peptide

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