製品の概要

  • 製品名Anti-Brd4 antibody
    Brd4 一次抗体 製品一覧
  • 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

Anti-Brd4 antibody 画像

  • 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).

     

    Secondary antibody - anti-mouse HRP (ab131366)

    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.

Anti-Brd4 antibody (ab75898) 使用論文

This product has been referenced in:
  • Baratta MG  et al. An in-tumor genetic screen reveals that the BET bromodomain protein, BRD4, is a potential therapeutic target in ovarian carcinoma. Proc Natl Acad Sci U S A 112:232-7 (2015). WB ; Human . Read more (PubMed: 25535366) »
  • Knoechel B  et al. An epigenetic mechanism of resistance to targeted therapy in T cell acute lymphoblastic leukemia. Nat Genet 46:364-70 (2014). WB ; Human . Read more (PubMed: 24584072) »

See all 6 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

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...

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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...

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