Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5150(2)] to Brd4 - BSA and Azide free
  • Suitable for: ICC/IF, Flow Cyt, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free
    See all Brd4 primary antibodies

  • Description

    Rabbit monoclonal [EPR5150(2)] to Brd4 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

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

  • Positive control

    • HAP1, HeLa, Caco2, T.T, RAW264.7 and NIH3T3 cell lysates; Human colon carcinoma tissue.

  • General notes

    ab182446 is the carrier-free version of ab128874. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

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

    ab182446 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    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

Images

  • Western blot - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    Western blot - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    All lanes : Anti-Brd4 antibody [EPR5150(2)] (ab128874) at 1/200 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : BRD4 knockout HAP1 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 152 kDa
    Observed band size: 220 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 (ab128874).

    Lanes 1 - 2: Merged signal (red and green). Green - ab128874 observed at 220 kDa. Red - loading control Mouse anti Vinculin observed at 125 kDa.

    ab128874 was shown to react with Brd4 in wild-type cells in Western blot with loss of signal observed in BRD4 knockout sample. Wild-type and BRD4 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab128874 and Mouse anti Vinculin overnight at 4 °C at a 1 in 200 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Flow Cytometry - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    Flow Cytometry - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)

    Flow cytometry analysis of SW480 (Human colorectal adenocarcinoma epithelial cell) cells labeling Brd4 (red) with purified ab128874 at a 1/50 dilution (10 µg/mL). Cells were fixed with 80% methanol and permeabilized with 0.1% Tween-20. A goat anti rabbit IgG (Alexa Fluor®488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128874).

  • Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)

    Immunocytochemistry/Immunofluorescence analysis of HepG2 (Human liver cell line) cells labeling Brd4 with purified ab128874 at 1/100 (Panel A). Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain(Panel B).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128874).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human brain tissue labeling Brd4 with purified ab128874 at 1/200. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. An undiluted Goat Anti-Rabbit IgG H&L (HRP)  was used as the secondary antibody at. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128874).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)

    Unpurified ab128874 at 1/100 dilution staining Brd4 in human colon carcinoma tissue by Immunohistochemistry.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128874).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    Immunocytochemistry/ Immunofluorescence - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446) This image is courtesy of an Abreview submitted by Kirk McManus.

    Unpurified ab128874 (1/500) staining Brd4 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells (green). Cells were fixed in paraformaldehyde, permeabilized in 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red).

    For further experimental details please refer to Abreview.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128874).

  • OI-RD Scanning - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    OI-RD Scanning - Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab128874).

  • Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)
    Anti-Brd4 antibody [EPR5150(2)] - BSA and Azide free (ab182446)

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