Diagenode

BRD4 polyclonal antibody

Antibody ChIP-seq grade icon
Catalog Number
Format
Price
C15410337
50 µg
$305.00
  Bulk order
Other format

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.

LotA2710P
Concentration2.6 µg/µl
Species reactivityHuman
TypePolyclonal
PurityAffinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.
HostRabbit
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 * 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

    ChIP

    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.ChIP-seq A
    B.ChIP-seq B
    C.ChIP-seq C
    D.ChIP-seq D

    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).

    ELISA

    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.

    WB

    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.

  •  Applications
    ChIP-seq
    Chromatin Immunoprecipitation (ChIP) coupled with high-throughput massively parallel sequencing as a detection method (ChIP-seq) has become one of the primary methods for epigenomics researchers, namely to investigate protein-DNA interaction on ... Read more
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    WB
    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
  •  Documents
    BRD4 polyclonal antibody DATASHEET
    Polyclonal antibody raised in rabbit against human BRD4 (Bromodomain Containing 4), using two KLH...
    Download
  •  Publications

    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!

Events

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    Nantes
    Oct 16-Oct 17, 2018
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    San Diego
    Oct 16-Oct 20, 2018
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