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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ubiquitin E3 Enzymes RING Finger E3 Ligase

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

Price and availability

367 686 ₸

Availability

Order now and get it on Tuesday March 02, 2021

Anti-BRD2 antibody [EPR7642] - ChIP Grade (ab139690)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

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.

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

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)

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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