Validated using a knockout cell line
Recombinant
RabMAb

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

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
    Suitable for: ICC/IF, Flow Cyt, WB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Brd4 aa 150-250. The exact sequence is proprietary.

  • Positive control
    • HeLa, Caco2, T.T, RAW264.7 and NIH3T3 cell lysates; Human colon carcinoma tissue.
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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 ab182446 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
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa).
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Target

  • Function
    Plays a role in a process governing chromosomal dynamics during mitosis.
  • Tissue specificity
    Ubiquitously expressed.
  • Involvement in disease
    Note=A chromosomal aberration involving BRD4 is found in a rare, aggressive, and lethal carcinoma arising in midline organs of young people. Translocation t(15;19)(q14;p13) with NUT which produces a BRD4-NUT fusion protein.
  • Sequence similarities
    Contains 2 bromo domains.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Brd4 antibody
    • BRD4-NUT FUSION antibody
    • BRD4-NUT fusion oncoprotein antibody
    • BRD4_HUMAN antibody
    • Bromodomain containing 4 antibody
    • bromodomain containing protein 4 antibody
    • Bromodomain-containing protein 4 antibody
    • CAP antibody
    • chromosome associated protein antibody
    • HUNK1 antibody
    • HUNKI antibody
    • MCAP antibody
    • Mitotic chromosome-associated protein antibody
    • Protein HUNK1 antibody
    see all

Images

  • 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 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) 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).

  • ab128874 at 1000 µg immunoprecipitating Brd4 in HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate. Sample was incubated with unpurified primary antibody (50 mM Tris dilution buffer) and protein G for 16 hours at 4°C.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

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

  • 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).

  • 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).

  • 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).

References

ab182446 has not yet been referenced specifically in any publications.

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