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RecombinantRabMAb

Recombinant Anti-Brd4 antibody [EPR25425-23] (ab289886)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR25425-23] (ab289886)
  • Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR25425-23] (ab289886)
  • Flow Cytometry (Intracellular) - Anti-Brd4 antibody [EPR25425-23] (ab289886)
  • Western blot - Anti-Brd4 antibody [EPR25425-23] (ab289886)
  • Flow Cytometry (Intracellular) - Anti-Brd4 antibody [EPR25425-23] (ab289886)
  • Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR25425-23] (ab289886)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR25425-23] (ab289886)
  • Western blot - Anti-Brd4 antibody [EPR25425-23] (ab289886)
  • Anti-Brd4 antibody [EPR25425-23] (ab289886)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR25425-23] to Brd4
  • Suitable for: ICC/IF, Flow Cyt (Intra), IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Brd4 antibody [EPR25425-23]
    See all Brd4 primary antibodies

  • Description

    Rabbit monoclonal [EPR25425-23] to Brd4

  • Host species

    Rabbit

  • Specificity

    This antibody does not react with Human species for IHC.

  • Tested applications

    Suitable for: ICC/IF, Flow Cyt (Intra), IHC-P, WBmore details
    Unsuitable for: ChIP or IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa, HEK-293T, NIH/3T3 and PC-12 whole cell lysates; Mouse testis tissue lysate; Rat testis tissue lysate. IHC-P: Mouse testis tissue; rat testis tissue. Flow cyt: HeLa and NIH/3T3 cells. ICC/IF: HeLa and NIH/3T3 cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab289886 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF
1/50.
Flow Cyt (Intra)
1/500.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 200 kDa (predicted molecular weight: 152 kDa).
Notes
ICC/IF
1/50.
Flow Cyt (Intra)
1/500.
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 200 kDa (predicted molecular weight: 152 kDa).
Application notes
Is unsuitable for ChIP or IP.

Target

  • Function

    Plays a role in a process governing chromosomal dynamics during mitosis.

  • Tissue specificity

    Ubiquitously expressed.

  • Involvement in disease

    Note=A chromosomal aberration involving BRD4 is found in a rare, aggressive, and lethal carcinoma arising in midline organs of young people. Translocation t(15;19)(q14;p13) with NUT which produces a BRD4-NUT fusion protein.

  • Sequence similarities

    Contains 2 bromo domains.

  • Cellular localization

    Nucleus.
  • Information by UniProt

  • Database links

    • Alternative names

      • Brd4 antibody
      • BRD4-NUT FUSION antibody
      • BRD4-NUT fusion oncoprotein antibody
      • BRD4_HUMAN antibody
      • Bromodomain containing 4 antibody
      • bromodomain containing protein 4 antibody
      • Bromodomain-containing protein 4 antibody
      • CAP antibody
      • chromosome associated protein antibody
      • HUNK1 antibody
      • HUNKI antibody
      • MCAP antibody
      • Mitotic chromosome-associated protein antibody
      • Protein HUNK1 antibody
      see all

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR25425-23] (ab289886)

      Immunohistochemical analysis of paraffin-embedded Rat testis tissue labelling Brd4 with ab289886 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nucleus staining was found in rat testis. The section was incubated with ab289886 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR25425-23] (ab289886)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labeling Brd4 with ab289886 at 1/50 dilution, followed by ab150081 Goat Anti-Rbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubµle Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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 dilution.

    • Flow Cytometry (Intracellular) - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Flow Cytometry (Intracellular) - Anti-Brd4 antibody [EPR25425-23] (ab289886)

      Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling Brd4 with ab289886 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled 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.

    • Western blot - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Western blot - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      All lanes : Anti-Brd4 antibody [EPR25425-23] (ab289886) at 1/1000 dilution

      Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : 293T (human embryonic kidney epithelial cell) whole cell lysate
      Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
      Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

      Predicted band size: 152 kDa
      Observed band size: 200 kDa why is the actual band size different from the predicted?



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

      Lysates were prepared from fresh material and used for Western blotting immediately to minimize protein degradation.

      Exposure time: 48 seconds.

       

    • Flow Cytometry (Intracellular) - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Flow Cytometry (Intracellular) - Anti-Brd4 antibody [EPR25425-23] (ab289886)

      Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling Brd4 with ab289886 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR25425-23] (ab289886)

      Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa cells labeling Brd4 with ab289886 at 1/50 dilution, followed by ab150081 Goat Anti-Rbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 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 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR25425-23] (ab289886)

      Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling Brd4 with ab289886 at 1/2000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining was found in mouse testis (PMID:28076398). The section was incubated with ab289886 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Western blot - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Western blot - Anti-Brd4 antibody [EPR25425-23] (ab289886)
      All lanes : Anti-Brd4 antibody [EPR25425-23] (ab289886) at 1/1000 dilution

      Lane 1 : Mouse testis tissue lysate
      Lane 2 : Rat testis tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

      Predicted band size: 152 kDa
      Observed band size: 200 kDa why is the actual band size different from the predicted?



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

      This blot was developed using a higher sensitivity ECL substrate.

      Exposure time: 180 seconds

       

    • Anti-Brd4 antibody [EPR25425-23] (ab289886)
      Anti-Brd4 antibody [EPR25425-23] (ab289886)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab289886? Please let us know so that we can cite the reference in this datasheet.

    ab289886 has not yet been referenced specifically in any publications.

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