Overview

  • Product nameAnti-Brd4 antibody [EPR5150(2)]
    See all Brd4 primary antibodies
  • Description
    Rabbit monoclonal [EPR5150(2)] to Brd4
  • Tested applicationsWB, IP, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Brd4 aa 150-250.
    Database link: O60885

  • Positive control
    • HeLa, Caco2, T.T, RAW264.7 and NIH3T3 cell lysates; HepG2 cells, Human colon carcinoma tissue and Human brain.
  • General notes

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

     Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

     

Properties

Applications

Our Abpromise guarantee covers the use of ab128874 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
WB 1/1000. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa).

For unpurified use at 1/200

IP Use at an assay dependent concentration.

For unpurified use at 1ug/ml of lysate

IHC-P 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

For unpurified use at 1/40

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

ICC/IF 1/200.

For unpurified use at 1/40

  • Application notesIs unsuitable for Flow Cyt.
  • Target

    • FunctionPlays a role in a process governing chromosomal dynamics during mitosis.
    • Tissue specificityUbiquitously expressed.
    • Involvement in diseaseNote=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 similaritiesContains 2 bromo domains.
    • Cellular localizationNucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • Brd4 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
      • Protein HUNK1 antibody
      see all

    Anti-Brd4 antibody [EPR5150(2)] images

    • Immunocytochemistry/Immunofluorescence analysis of HepG2 (Human liver cell line) cells labelling Brd4 with purified ab128874 at 1/100 (A). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counter stain(B).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human brain tissue labelling Brd4 with purified ab128874 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. An undiluted Goat Anti-Rabbit IgG H&L (HRP)  was used as the secondary antibody at. Counter stained with Hematoxylin.

    • Anti-Brd4 antibody [EPR5150(2)] (ab128874) at 1/1000 dilution (Purified) + NIH/3T3 Cell Lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size : 152 kDa
      Observed band size : 152 kDa
    • Anti-Brd4 antibody [EPR5150(2)] (ab128874) at 1/1000 dilution (purified) + HeLa at 10 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size : 152 kDa
      Observed band size : 152 kDa

      Blocking and Diluting buffer and concentration: 5% NFDM/TBST

    • Anti-Brd4 antibody [EPR5150(2)] (ab128874) at 1/200 dilution (unpurified) + HeLa at 10 µg/ml

      Secondary
      Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size : 152 kDa
      Observed band size : 152 kDa

      Blocking and Diluting buffer and concentration: 5% NFDM/TBST

    • Unpurified ab128874 at 1/100 dilution staining Brd4 in Human colon carcinoma tissue by Immunohistochemistry.

    • Unpurified ab128874 (1/500) staining Brd4 in Hela cells (green). Cells were fixed in paraformaldehyde, permeabilised in 0.5% Triton X100/PBS and counter stained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

      See Abreview

    • ab128874 Immunoprecipitating Brd4 in human HEK293 whole cell lysate. 1000µg of cell lysate was incubated with unpurified primary antibody (1 µg/ml in 50 mM TRIS) and matrix (Protein G) for 16 hours at 4°C. For western blotting ab131366 (anti-mouse HRP, 1/10000) was used to confirm successful immunoprecipation.

      See Abreview

    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References for Anti-Brd4 antibody [EPR5150(2)] (ab128874)

    This product has been referenced in:
    • Sahai V  et al. BET bromodomain inhibitors block growth of pancreatic cancer cells in three-dimensional collagen. Mol Cancer Ther 13:1907-17 (2014). Human . Read more (PubMed: 24807963) »

    See 1 Publication for this product

    Product Wall

    Application Western blot
    Loading amount 30 µg
    Gel Running Conditions Non-reduced Non-Denaturing (Native)
    Sample Mouse Cell lysate - whole cell (pancreatic cancer)
    Specification pancreatic cancer
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Username

    Ms. Jamie Plevy

    Verified customer

    Submitted Sep 26 2014

    Application Immunoprecipitation
    Immuno-precipitation step Protein G
    Sample Human Cell lysate - whole cell (HEK293)
    Specification HEK293
    Total protein in input 1000 µg
    Username

    Abcam user community

    Verified customer

    Submitted Apr 25 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
    Loading amount 15 µg
    Specification mouse embryonic fibroblasts
    Gel Running Conditions Reduced Denaturing
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 23 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Cell lysate - whole cell (pancreatic tumor)
    Loading amount 25 µg
    Specification pancreatic tumor
    Gel Running Conditions Reduced Denaturing
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Nov 07 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (pancreatic cancer cell line)
    Loading amount 25 µg
    Specification pancreatic cancer cell line
    Gel Running Conditions Reduced Denaturing
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Nov 07 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunocytochemistry/ Immunofluorescence
    Sample Human Cell (HeLa)
    Specification HeLa
    Fixative Paraformaldehyde
    Permeabilization Yes - 0.5% TritonX100 in PBS
    Username

    Dr. Kirk McManus

    Verified customer

    Submitted Oct 19 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Human Tissue sections (endometrial adenocarcinoma)
    Specification endometrial adenocarcinoma
    Fixative Paraformaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
    Permeabilization Yes - PBST
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 08 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (endometrial adenocarcinoma)
    Specification endometrial adenocarcinoma
    Fixative Paraformaldehyde
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate
    Permeabilization Yes - PBST
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Oct 08 2012

    Thank you very much for your call last week and for your patience while I have been in touch with the lab regarding ab128874.

    The immunogen is a peptide taken from between amino acids 175 and 200 of human Brd4.

    I hope this informati...

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"