Overview

  • Product nameAnti-Brd4 antibodySee all Brd4 primary antibodies ...
  • Description
    Rabbit polyclonal to Brd4
  • Tested applicationsICC/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.

    (Peptide available as ab85811.)

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

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).Can be blocked with Brd4 peptide (ab85811).

Target

  • FunctionPlays a role in a process governing chromosomal dynamics during mitosis.
  • Tissue specificityUbiquitously expressed.
  • Involvement in diseaseNote=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 similaritiesContains 2 bromo domains.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Brd4 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
    • Protein HUNK1 antibody
    see all

Anti-Brd4 antibody images

  • ab75898 Immunoprecipitating Brd4 in human HEK293 whole cell lysate. 1000µg of cell lysate was incubated with primary antibody (1 µg/mg lysate in 50mM TRIS) and matrix (Protein G) for 16 hours at 4°C. For western blotting ab131366 (1/10000) was used to confirm successful immunoprecipation.

    Detects band around 150 kDa (predicted size) also additional band around 250 kDa (possibly modified Brd4).

    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 : Liver (Human) Tissue Lysate - adult normal tissue (ab29889)

    Lysates/proteins at 10 µg/ml per lane.

    Secondary
    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 for Anti-Brd4 antibody (ab75898)

This product has been referenced in:
  • Liu P  et al. Release of positive transcription elongation factor b (P-TEFb) from 7SK small nuclear ribonucleoprotein (snRNP) activates hexamethylene bisacetamide-inducible protein (HEXIM1) transcription. J Biol Chem 289:9918-25 (2014). WB ; Human . Read more (PubMed: 24515107) »
  • Hussong M  et al. The bromodomain protein BRD4 regulates the KEAP1/NRF2-dependent oxidative stress response. Cell Death Dis 5:e1195 (2014). Human . Read more (PubMed: 24763052) »

See all 4 Publications for this product

Product Wall

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

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
Username

Abcam user community

Verified customer

Submitted Jan 29 2013

Thank you for taking time to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring...

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

Dr. Kirk McManus

Verified customer

Submitted Feb 03 2012

Thank you for your inquiry. I am very sorry to hear that you are having problem with this antibody. As I understand from your e-mail, there is some problem with co-IP. Unfortunately, as the datasheet indicates this product has been tested in IC...

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