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BRD4 polyclonal antibody

50 µg
  Bulk order

Polyclonal antibody raised in rabbit against human BRD4 (Bromodomain Containing 4), using two KLH-conjugated synthetic peptides from the N-terminal and central part of the protein, respectively.

Concentration2.6 µg/µl
Species reactivityHuman
PurityAffinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.
Storage ConditionsStore at -20°C; for long storage, store at -80°C. Avoid multiple freeze-thaw cycles.
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution/amount References
ChIP/ChIP-seq * 2 µg/ChIP Fig 1, 2
ELISA 1:10,000 Fig 3
Western blotting 1:1,000 Fig 4

*Please note that the optimal antibody amount per IP should be determined by the end-user. We recommend testing 1-5 µg per IP.

  • Validation Data

    BRD4 Antibody ChIP Grade

    Figure 1. ChIP results obtained with the Diagenode antibody directed against BRD4
    ChIP was performed with the Diagenode antibody against BRD4 (Cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells. An antibody titration consisting of 1, 2, 5 and 10 µg per ChIP experiment was analysed. IgG (2 µg/IP) was used as negative IP control. QPCR was performed with primers specific for the EIF4A2 and ACTB promoters, used as positive controls, and for the MYOD1 gene and Sat2 satellite repeat, used as negative controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    A. BRD4 Antibody ChIP-seq Grade
    B. BRD4 Antibody for ChIP-seq
    C. BRD4 Antibody for ChIP-seq assay
    D. BRD4 Antibody validated in ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against BRD4
    ChIP was performed with 2 µg of the Diagenode antibody against BRD4 (cat. No. C15410337) on sheared chromatin from 4,000,000 K562 cells using the “iDeal ChIP-seq” kit as described above. The IP'd DNA was subsequently analysed on an Illumina HiSeq 2000. Library preparation, cluster generation and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the signal distribution along the complete sequence and a 300 kb region of the human X-chromosome (figures 2A and B), and in two genomic rtegions surrounding the ACTB and EIF4A2 positive control genes (figure 2C and D).

    BRD4 Antibody ELISA Validation

    Figure 3. Determination of the antibody titer
    To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against BRD4 (Cat. No. C15410337). The plates were coated with the peptides used for immunization of the rabbit. By plotting the absorbance against the antibody dilution (Figure 2), the titer of the antibody was estimated to be 1:290,000.

    BRD4 Antibody validated in Western Blot

    Figure 4. Western blot analysis using the Diagenode antibody directed against BRD4
    Whole cell extracts (25 µg) from HeLa cells were analysed by Western blot using the Diagenode antibody against BRD4 (Cat. No. C15410337) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

  • Target Description

    BRD4 (UniProt/Swiss-Prot entry O60885) is a chromatin reader protein that binds acetylated histones. It remains associated with acetylated chromatin throughout the entire cell cycle and provides epigenetic memory for gene transcription by preserving an acetylated chromatin status. As such, it plays a key role in the transmission of epigenetic memory across cell divisions. BRD4 promotes phosphorylation of Ser-2 of the C-terminal domain (CTD) of RNA polymerase II and plays a key role in regulating the transcription of signal-inducible genes. It has been implicated in a translocation of chromosome 19 which causes an upper respiratory tract carcinoma.

  •  実験手法
    検出方法(ChIP-seq)としてハイスループットシーケンシングと結合したクロマチン免疫沈降(ChIP)は、今やエピゲノム研究、すなわちゲノム規模でのタンパク質-DNA相互作用の研究において主要な方法の1つされています。また、この技術は現在、細胞分化、腫瘍抑制遺伝子のサイレンシング、および遺伝子発現に対するヒストン修飾の効果を含む様々なライフサイエンス分野で使用されています。 ChIP-seqワークフロー クロマチン調製 : DNAへのヒストンまたは転... Read more
    Enzyme-linked immunosorbent assay. Read more
    Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies. Learn more about: Load... Read more
  •  資料
    BRD4 polyclonal antibody DATASHEET
    Polyclonal antibody raised in rabbit against human BRD4 (Bromodomain Containing 4), using two KLH...
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: BRD4 polyclonal antibody (Diagenode Cat# C15410337 Lot# A2710P). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    DeltaNp63-dependent super enhancers define molecular identity in pancreatic cancer by an interconnected transcription factor network.
    Hamdan FH, Johnsen SA
    Molecular subtyping of cancer offers tremendous promise for the optimization of a precision oncology approach to anticancer therapy. Recent advances in pancreatic cancer research uncovered various molecular subtypes with tumors expressing a squamous/basal-like gene expression signature displaying a worse prognosis. ...

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