Diagenode

NFKB p65 polyclonal antibody - Classic

カタログ番号
フォーマット
価格
C15310256
100 μl
$345.00
  Bulk order
ブローカー情報

Alternative names: RELA, NFKB3, p65

Polyclonal antibody raised in rabbit against NFKB p65 (Rel A), using a KLH-conjugated synthetic peptide.

Lot24468
Concentrationnot determined
Species reactivityHuman, mouse, rat
TypePolyclonal
PurityWhole antiserum
HostRabbit
PrecautionsThis product is for research use only. Not for use in diagnostic or therapeutic procedures.
Applications Suggested dilution References
ChIP/ChIP-seq * 1 µl/IP Fig 1, 2
Western Blotting 1:2,000 - 1:5,000 Fig 3
ELISA 1:5,000
Immunohistochemistry 1:500 - 1:2,000 Fig 4
Gel Shift 1:500

* 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 NFkB p65.
    ChIP assays were performed using human HeLa cells, treated with TNFalpha, the Diagenode antibody against NFkB p65 (Cat. No. C15310256) and optimized PCR primer sets for qPCR. ChIP was performed with the “iDeal ChIP-seq” kit (Cat. No. C01010055), using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2.5 and 5 µl per ChIP experiment was analysed. IgG (1 µg/IP) was used as negative IP control. QPCR was performed with primers for the NFKBIA and CCL20 genes, used as positive controls, and for TSH2B, used as negative control. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    ChIP-seq

    ChIP-seq

    ChIP-seq

    ChIP-seq

    Figure 2. ChIP-seq results obtained with the Diagenode antibody directed against NFkB p65
    ChIP was performed on sheared chromatin from 4 million HeLa cells using 1 µg of the Diagenode antibody against NFkB p65 (Cat. No. C15310256) as described above. The IP’d DNA was subsequently analysed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer’s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure 2 shows the enrichment along the complete sequence and a 2 Mb region of human chromosome 2 (fig 2A and B), and in a two genomic regions surrounding the NFKBIA and CCL20 positive control genes.

    WB

    Figure 3. NFKB p65 antibody western blot results
    Whole cell extracts from HeLa cells (35 µg) were analysed by Western blot using the Diagenode antibody against NFkB p65 (Cat. No. C15310256) diluted 1:5,000. The position of the protein of interest is indicated on the right (expected size: 65 kDa); the marker (in kDa) is shown on the left.

    Immunohistochemistry

    Figure 4. NFKB p65 antibody Immunohistochemistry results Formalin fixed paraffin embedded lymphocytes and germinal center cells of the tonsil were stained with the Diagenode antibody against NFkB p65 (Cat. No. C15310256) diluted 1:400 followed by a peroxidase labelled goat anti-rabbit secondary antibody. Figure 4 shows moderate positive nuclear or cytoplasmic staining.

  •  実験手法
    ELISA
    Enzyme-linked immunosorbent assay. Read more
    IHC
    Immunohistochemistry 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
    ChIP-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  •  資料
    Datasheet NFKBp65 C15310256 DATASHEET
    Datasheet description
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    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
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    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
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  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: NFKB p65 polyclonal antibody - Classic (Diagenode Cat# C15310256 Lot# 24468). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    RRAD, IL4I1, CDKN1A, and SERPINE1 genes are potentially co-regulated by NF-κB and p53 transcription factors in cells exposed to high doses of ionizing radiation.
    Szołtysek K, Janus P, Zając G, Stokowy T, Walaszczyk A, Widłak W, Wojtaś B, Gielniewski B, Cockell S, Perkins ND, Kimmel M, Widlak P
    BACKGROUND: The cellular response to ionizing radiation involves activation of p53-dependent pathways and activation of the atypical NF-κB pathway. The crosstalk between these two transcriptional networks include (co)regulation of common gene targets. Here we looked for novel genes potentially (co)regulated by...

    Pro-inflammatory cytokine and high doses of ionizing radiation have similar effects on the expression of NF-kappaB-dependent genes.
    Janus P, Szołtysek K, Zając G, Stokowy T, Walaszczyk A, Widłak W, Wojtaś B, Gielniewski B, Iwanaszko M, Braun R, Cockell S, Perkins ND, Kimmel M, Widlak P
    The NF-κB transcription factors are activated via diverse molecular mechanisms in response to various types of stimuli. A plethora of functions associated with specific sets of target genes could be regulated differentially by this factor, affecting cellular response to stress including an anticancer treatment...

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