Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)

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 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), ChIP, ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-BRD2 antibody [EPR7642] - BSA and Azide free
    See all BRD2 primary antibodies

  • Description

    Rabbit monoclonal [EPR7642] to BRD2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), ChIP, ICC/IF, WB, IHC-Pmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

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

  • General notes

    ab222393 is the carrier-free version of ab139690.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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

Images

  • Western blot - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    Western blot - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    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?



    This data was developed using the same antibody clone in a different buffer formulation (ab139690).

    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] - BSA and Azide free (ab222393)
    ChIP - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)

    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
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab139690).

  • Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab139690).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab139690).

    Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    Immunocytochemistry/ Immunofluorescence - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    Immunofluorescence analysis of HeLa cells labeling BRD2, with ab139690 at 1/250 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab139690).

  • Flow Cytometry - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    Flow Cytometry - Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab139690).

  • Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)
    Anti-BRD2 antibody [EPR7642] - BSA and Azide free (ab222393)

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