製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to Brd4
  • アプリケーション
    適用あり: ICC/IF, IP, WBmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Horse, Dog
  • 免疫原

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

    (Peptide available as ab85811.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in Human, Mouse and Rat Liver Tissue Lysates.
  • 特記事項

    "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."

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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).

ターゲット情報

  • 機能
    Plays a role in a process governing chromosomal dynamics during mitosis.
  • 組織特異性
    Ubiquitously expressed.
  • 関連疾患
    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.
  • 配列類似性
    Contains 2 bromo domains.
  • 細胞内局在
    Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像



  • Predicted band size : 152 kDa

    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, ab131366 was used as the secondary antibody at 1/10000 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
    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.

参考文献

This product has been referenced in:
  • Col E  et al. Bromodomain factors of BET family are new essential actors of pericentric heterochromatin transcriptional activation in response to heat shock. Sci Rep 7:5418 (2017). Read more (PubMed: 28710461) »
  • Boi M  et al. Therapeutic efficacy of the bromodomain inhibitor OTX015/MK-8628 in ALK-positive anaplastic large cell lymphoma: an alternative modality to overcome resistant phenotypes. Oncotarget 7:79637-79653 (2016). WB . Read more (PubMed: 27793034) »

See all 14 Publications for this product

レビューと Q&A

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

投稿 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
Username

Abcam user community

Verified customer

投稿 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

投稿 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...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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

投稿 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...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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