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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ubiquitin E3 Enzymes RING Finger E3 Ligase
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)

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  • SDS
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Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • ChIP - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • Flow Cytometry - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR7642] to BRD2 - ChIP Grade
  • Suitable for: WB, Flow Cyt, ICC/IF, IHC-P, ChIP
  • Knockout validated
  • Reacts with: Human

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Overview

  • Product name

    Anti-BRD2 antibody [EPR7642] - ChIP Grade
    See all BRD2 primary antibodies
  • Description

    Rabbit monoclonal [EPR7642] to BRD2 - ChIP Grade
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    ChIP
    Human
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

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

  • Positive control

    • WB: HEK293T, Jurkat, MOLT4, NCCIT and HeLa whole cell lysate (ab150035). ICC/IF: HeLa and wild-type HAP1 cells. IHC-P: Human testis tissue. ChIP: Nuclear extract from LNCaP 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20ºC.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EPR7642
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ubiquitin E3 Enzymes
    • RING Finger E3 Ligase
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Developmental Families
    • Other
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Bromodomain containing
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies
    • Epigenetics and Nuclear Signaling
    • Bromodomains

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-BRD2 antibody [EPR7642] (ab197865)
    • Alexa Fluor® 647 Anti-BRD2 antibody [EPR7642] (ab197866)
    • HRP Anti-BRD2 antibody [EPR7642] (ab198536)
    • Alexa Fluor® 555 Anti-BRD2 antibody [EPR7642] (ab213347)
    • Alexa Fluor® 594 Anti-BRD2 antibody [EPR7642] (ab215507)
    • Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human BRD2 knockout HEK-293T cell line (ab267265)
  • KO cell lysates

    • Human BRD2 knockout HEK-293T cell lysate (ab257191)
  • KO cell pellets

    • Human BRD2 knockout HEK293T cell pellet (ab278860)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab7899)
    • MOLT-4 whole cell lysate (ab7912)
  • Recombinant Protein

    • Recombinant Human BRD2 protein (ab187435)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ChIP
Human
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
WB
Human
Application Abreviews Notes
WB (2)
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 (2)
1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ChIP
Use at an assay dependent concentration.
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.
ChIP
Use at an assay dependent concentration.
  • 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.
    • Target information above from: UniProt accession P25440 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 6046 Human
      • Omim: 601540 Human
      • SwissProt: P25440 Human
      • Unigene: 75243 Human
      • 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

      • Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        All lanes : Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690) at 1/1000 dilution

        Lane 1 : Wild-type HEK293T cell lysate
        Lane 2 : BRD2 knockout HEK293T cell lysate
        Lane 3 : HeLa cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

        Predicted band size: 88 kDa
        Observed band size: 110 kDa why is the actual band size different from the predicted?



        Lanes 1-3: Merged signal (red and green). Green - ab139690 observed at 110 kDa. Red - loading control ab8245 observed at 36 kDa. 

         ab139690 Anti-BRD2 antibody [EPR7642]  was shown to specifically react with BRD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267265 (knockout cell lysate ab257191) was used.  Wild-type and BRD2 knockout samples were subjected to SDS-PAGE.  ab139690 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.

      • ChIP - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        ChIP - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Chromatin was prepared from LNCaP cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with formaldehyde for 10 minutes.
        The ChIP was performed with 25 µg of chromatin, 5 µg of ab139690 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
        Primers and probes are located in the first kb of the transcribed region.
        *http://www.abcam.com/resources?keywords=X%20ChIP%20protocol
      • Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)

        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.

      • Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Western blot - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        All lanes : Anti-BRD2 antibody [EPR7642] - ChIP Grade (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

      • Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)

        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).

      • Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Immunofluorescence analysis of HeLa cells labeling BRD2, with ab139690 at 1/250 dilution.
      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)

        Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling BRD2 with ab139690 at 1/250 dilution.

        Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

      • Flow Cytometry - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Flow Cytometry - Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        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.
      • Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
        Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)

      Protocols

      • Flow cytometry protocols
      • Immunohistochemistry protocols
      • Immunocytochemistry & immunofluorescence protocols
      • Western blot protocols

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (18)

      Publishing research using ab139690? Please let us know so that we can cite the reference in this datasheet.

      ab139690 has been referenced in 18 publications.

      • Wakita M  et al. A BET family protein degrader provokes senolysis by targeting NHEJ and autophagy in senescent cells. Nat Commun 11:1935 (2020). PubMed: 32321921
      • Yang C  et al. FBP1 binds to the bromodomain of BRD4 to inhibit pancreatic cancer progression. Am J Cancer Res 10:523-535 (2020). PubMed: 32195024
      • Wang H  et al. BRD4 contributes to LPS-induced macrophage senescence and promotes progression of atherosclerosis-associated lipid uptake. Aging (Albany NY) 12:9240-9259 (2020). PubMed: 32392533
      • Tian XP  et al. BRD2 induces drug resistance through activation of the RasGRP1/Ras/ERK signaling pathway in adult T-cell lymphoblastic lymphoma. Cancer Commun (Lond) 40:245-259 (2020). PubMed: 32459053
      • Wessel SR  et al. Functional Analysis of the Replication Fork Proteome Identifies BET Proteins as PCNA Regulators. Cell Rep 28:3497-3509.e4 (2019). PubMed: 31553917
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-4 of 4 Abreviews or Q&A

      Western blot abreview for Anti-BRD2 antibody [EPR7642] - ChIP Grade

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Cell lysate - whole cell (mouse neural stem cells)
      Gel Running Conditions
      Reduced Denaturing (4-12%)
      Loading amount
      25 µg
      Specification
      mouse neural stem cells
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
      Read More

      Dr. Robert Kupp

      Verified customer

      Submitted Oct 19 2020

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BRD2 antibody [EPR7642]

      Good
      Abreviews
      Abreviews
      abreview image
      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
      Read More

      Abcam user community

      Verified customer

      Submitted Jun 20 2017

      Western blot abreview for Anti-BRD2 antibody [EPR7642] - ChIP Grade

      Excellent
      Abreviews
      Abreviews
      abreview image
      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
      Read More

      Abcam user community

      Verified customer

      Submitted Nov 20 2015

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BRD2 antibody [EPR7642] - ChIP Grade

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

      Abcam user community

      Verified customer

      Submitted Nov 17 2014

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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