Validated using a knockout cell line
Recombinant
RabMAb

Anti-BRD2 antibody [EPR7642] (ab139690)

Overview

  • Product name
    Anti-BRD2 antibody [EPR7642]
    See all BRD2 primary antibodies
  • Description
    Rabbit monoclonal [EPR7642] to BRD2
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IF, IHC-Pmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human BRD2 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: P25440

  • Positive control
    • WB: Jurkat, MOLT4, NCCIT and HeLa cell lysates. ICC/IF: HeLa and wild-type HAP1 cells. IHC-P: Human testis tissue.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab139690 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 - 1/10000. Predicted molecular weight: 88 kDa.
Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use a concentration of 0.5 µg/ml.
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      May play a role in spermatogenesis or folliculogenesis.
    • Sequence similarities
      Contains 2 bromo domains.
      Contains 1 ET domain.
    • Domain
      One bromodomain is sufficient for a partial interaction with histone H4 acetylated at 'Lys-13'.
    • Cellular localization
      Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • BRD 2 antibody
      • Brd2 antibody
      • BRD2_HUMAN antibody
      • Bromodomain containing 2 antibody
      • Bromodomain-containing protein 2 antibody
      • D6S113E antibody
      • DKFZp686N0336 antibody
      • Female sterile homeotic related gene 1 antibody
      • Female sterile homeotic related gene 1, mouse, homolog of antibody
      • FLJ31942 antibody
      • FSH antibody
      • FSRG1 antibody
      • KIAA9001 antibody
      • NAT antibody
      • O27.1.1 antibody
      • Really interesting new gene 3 protein antibody
      • RING3 antibody
      • RING3 PROTEIN antibody
      • RNF3 antibody
      see all

    Images

    • Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: BRD2 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: A431 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab139690observed at 110 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab139690 was shown to specifically react with BRD2 when BRD2 knockout samples were used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. ab139690 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • ab139690 staining Brd2 in wild-type HAP1 cells (top panel) and BRD2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab139690 at 0.5μg/ml and ab195889 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Overlay histogram showing HeLa cells stained with ab139690 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab139690, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • All lanes : Anti-BRD2 antibody [EPR7642] (ab139690) at 1/1000 dilution

      Lane 1 : MOLT4 cell lysate
      Lane 2 : Jurkat cell lysate
      Lane 3 : NCCIT cell lysate
      Lane 4 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Standard HRP labelled goat anti-rabbit at 1/2000 dilution

      Developed using the ECL technique.

      Predicted band size: 88 kDa

    • Immunofluorescence analysis of HeLa cells labeling BRD2, with ab139690 at 1/250 dilution.
    • Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling BRD2 with ab139690 at 1/250 dilution.

    References

    This product has been referenced in:
    • Vázquez R  et al. The bromodomain inhibitor OTX015 (MK-8628) exerts anti-tumor activity in triple-negative breast cancer models as single agent and in combination with everolimus. Oncotarget 8:7598-7613 (2017). WB . Read more (PubMed: 27935867) »
    • Zhao L  et al. Photoreceptor protection via blockade of BET epigenetic readers in a murine model of inherited retinal degeneration. J Neuroinflammation 14:14 (2017). Read more (PubMed: 28103888) »

    See all 5 Publications for this product

    Customer reviews and Q&As

    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 (Testis)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Ventana CC1 (EDTA pH8.5)
    Permeabilization
    No
    Specification
    Testis
    Blocking step
    Ventana blocking agent as blocking agent for 4 minute(s) · Concentration: 100% · Temperature: 36°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jun 20 2017

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (mouse B6 ESC, E13.5 neurosphere, E13.5 MEF, N2a, E)
    Gel Running Conditions
    Reduced Denaturing (8%)
    Loading amount
    60 µg
    Specification
    mouse B6 ESC, E13.5 neurosphere, E13.5 MEF, N2a, E
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Abcam user community

    Verified customer

    Submitted Nov 20 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Compact polymer kit used, no additional as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 20°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate Buffer, 6.0 pH
    Sample
    Human Tissue sections (Heart)
    Specification
    Heart
    Permeabilization
    Yes - Surfactant in washing reagent
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Nov 17 2014

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

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