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Recombinant

Recombinant Anti-Brd4 antibody [BL-149-2H5] (ab243862)

Reviews (1) Submit a question References (2)
Western blot - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
  • Immunoprecipitation - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
  • ChIP-sequencing - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
  • Western blot - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
  • Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
  • Flow Cytometry - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
  • Anti-Brd4 antibody [BL-149-2H5] (ab243862)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [BL-149-2H5] to Brd4
  • Suitable for: WB, ChIP-sequencing, ICC/IF, IP, Flow Cyt
  • Reacts with: Mouse, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Brd4 antibody [BL-149-2H5]
    See all Brd4 primary antibodies

  • Description

    Rabbit monoclonal [BL-149-2H5] to Brd4

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, ChIP-sequencing, ICC/IF, IP, Flow Cytmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Synthetic peptide within Human Brd4 aa 1312-1362. The exact sequence is proprietary. NP_490597.1 and Gene ID 23476.
    Database link: O60885

  • Positive control

    • WB: HeLa, HEK293T, Jurkat, MDA-MB-231, A-549, SW620, SK-MEL-28, BW5147.3 whole cell lysate. IP: HeLa whole cell lysate. ChIP: Mia PaCa-2 cells. ICC/IF: HeLa cells. Flow Cyt: KG-1 cells.

  • General notes

    This product is sold under License from Bethyl Laboratories, Inc.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab243862 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.
ChIP-sequencing
Use a concentration of 10-40 - 30 µl/chromatin.
ICC/IF
1/25 - 1/250.

Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells.
IP
Use at an assay dependent concentration.

Use 5-10µl/0.5mg lysate.
Flow Cyt
Use 1µl for 106 cells.
Notes
WB
1/1000.
ChIP-sequencing
Use a concentration of 10-40 - 30 µl/chromatin.
ICC/IF
1/25 - 1/250.

Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells.
IP
Use at an assay dependent concentration.

Use 5-10µl/0.5mg lysate.
Flow Cyt
Use 1µl for 106 cells.

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

    • Western blot - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Western blot - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      All lanes : Anti-Brd4 antibody [BL-149-2H5] (ab243862) at 1/1000 dilution

      Lane 1 : HeLa whole cell lysate
      Lane 2 : HEK293T whole cell lysate
      Lane 3 : Jurkat whole cell lysate
      Lane 4 : MDA-MB-231 whole cell lysate
      Lane 5 : A-549 whole cell lysate
      Lane 6 : SW620 whole cell lysate
      Lane 7 : SK-MEL-28 whole cell lysate

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat anti-rabbit IgG

      Exposure time: 3 seconds


      Detection: Chemiluminescence.

      Lysates prepared using NETN lysis buffer.

    • Immunoprecipitation - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Immunoprecipitation - Anti-Brd4 antibody [BL-149-2H5] (ab243862)

      brd4 was immunoprecipitated from 1 mg HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate with ab243862 at 20 µL/mg lysate. Western blot was performed on the immunoprecipitate using ab243862 at 1/1000 dilution.

      Lane 1: ab243862 IP in HeLa cell lysate.

      Lane 2: Contol IgG in HeLa cell lysate.

    • ChIP-sequencing - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      ChIP-sequencing - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Localization of human BRD4 binding sites in immunoprecipitates from Mia PaCa-2 cells by ChIP-Seq using ab243862.
    • Western blot - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Western blot - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Anti-Brd4 antibody [BL-149-2H5] (ab243862) at 1/1000 dilution + BW5147.3 (Mouse T cell lymphoma cell line from thymus) whole cell lysate at 50 µg

      Secondary
      HRP-conjugated goat anti-rabbit IgG antibody


      Chemiluminescence with an exposure time of 3 seconds.

    • Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Formaldehyde-fixed HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling brd4 using ab243862 at 1/250 dilution in ICC/IF analysis. DyLight® 594-conjugated goat anti-rabbit IgG was used as the secondary antibody.
    • Flow Cytometry - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Flow Cytometry - Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Flow cytometric analysis of 4% formaldehyde and permeabilized with 90% methanol KG-1 (Human acute myeloid leukemia cell line from bone marrow) cells labelling BRD4 with ab243862 at 1µl/106 cells (Red shaded) compared with a Isotype control (unshaded). DyLight® 650-conjugated goat anti-rabbit IgG was used as the secondary antibody.
       
    • Anti-Brd4 antibody [BL-149-2H5] (ab243862)
      Anti-Brd4 antibody [BL-149-2H5] (ab243862)

    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 (2)

    Publishing research using ab243862? Please let us know so that we can cite the reference in this datasheet.

    ab243862 has been referenced in 2 publications.

    • Wu Z & Chen D MiR-129-5p Inactivates NF-κB Pathway to Block Rheumatoid Arthritis Development via Targeting BRD4. J Healthc Eng 2022:8330659 (2022). PubMed: 35494514
    • Zhao X  et al. Apigenin suppresses proliferation, invasion, and epithelial-mesenchymal transition of cervical carcinoma cells by regulation of miR-152/BRD4 axis. Kaohsiung J Med Sci 37:583-593 (2021). PubMed: 33611824

    Customer reviews and Q&As

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    ChIP abreview for Anti-Brd4 antibody [BL-149-2H5]

     Excellent
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Mouse Cell lysate - whole cell (LßT2 cells (mouse pituitary gonadotrope cell line))
    Negative control
    mSrd5a1 promoter, mMyc SuperEnhancer
    Specification
    LßT2 cells (mouse pituitary gonadotrope cell line)
    Detection step
    Real-time PCR
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: 1% formaldehyde
    Positive control
    mFshb regulatory region
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Oct 13 2021

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