Overview

  • Product name
  • Description
    Rabbit polyclonal to Brd4
  • Host species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Horse, Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1100 - 1200 of Human Brd4.

    Read Abcam's proprietary immunogen policy (Peptide available as ab85811.)

  • Positive control
    • This antibody gave a positive signal in Human, Mouse and Rat Liver Tissue Lysates.
  • General notes

    "Note: Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help."

Properties

Applications

Our Abpromise guarantee covers the use of ab75898 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 a concentration of 5 µg/ml.
IP Use at 1 µg/mg of lysate.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 152 kDa (predicted molecular weight: 152 kDa).

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

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Brd4 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Hu brain whole cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab75898 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

  • ab75898 at 1000µg immunoprecipitating Brd4 in HEK293 whole cell lysate. Sample was incubated with primary antibody (50mM 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.

    See Abreview

  • All lanes : Anti-Brd4 antibody (ab75898) at 1 µg/ml

    Lane 1 : Liver (Mouse) Tissue Lysate
    Lane 2 : Liver (Rat) Tissue Lysate
    Lane 3 : Human liver tissue lysate - total protein (ab29889)

    Lysates/proteins at 10 µg/ml per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP)

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 152 kDa
    Observed band size: 171 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 70 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 150 seconds
  • ab75898 (1/500) staining Brd4 in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilized with 0.5% Triton X-100/PBS and counterstained with DAPI in order to highlight the nucleus (red). For further experimental details please refer to Abreview.

    See Abreview

  • ICC/IF image of ab75898 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab75898, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and HepG2 cells at 5µg/ml.

References

This product has been referenced in:
  • Sui S  et al. Ferritinophagy is required for the induction of ferroptosis by the bromodomain protein BRD4 inhibitor (+)-JQ1 in cancer cells. Cell Death Dis 10:331 (2019). Read more (PubMed: 30988278) »
  • Wang C  et al. miR-204 enhances p27 mRNA stability by targeting Brd4 in head and neck squamous cell carcinoma. Oncol Lett 16:4179-4184 (2018). Read more (PubMed: 30250532) »
See all 20 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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1-4 of 4 Abreviews

Abreviews
Application
ChIP
Sample
Mouse Cell lysate - other (JB 6P+ cell)
Negative control
IgG
Specification
JB 6P+ cell
Detection step
Real-time PCR
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Positive control
DNA samples

Abcam user community

Verified customer

Submitted May 22 2017

Application
Immunoprecipitation
Immuno-precipitation step
Protein G
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Total protein in input
1000 µg

Abcam user community

Verified customer

Submitted May 08 2014

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse embryonic fibroblasts)
Loading amount
15 µg
Specification
mouse embryonic fibroblasts
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 29 2013

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton-X100 in PBS

Dr. Kirk Mcmanus

Verified customer

Submitted Feb 03 2012

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