NFKB p65 polyclonal antibody

Catalog Number
100 μl
  Bulk order

Alternative names: RELA, NFKB3, p65

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

Concentrationnot determined
Species reactivityHuman, mouse, rat
PurityWhole antiserum
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

    NFKB p65 Antibody ChIP Grade

    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).

    NFKB p65 Antibody ChIP-seq Grade

    NFKB p65 Antibody for ChIP-seq

    NFKB p65 Antibody for ChIP-seq assay

    NFKB p65 Antibody validated in 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.

    NFKB p65 Antibody validated in Western Blot

    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.

    NFKB p65 Antibody validated in 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.

  • Target Description

    This antibody recognizes NFKB p65 which is a component of NFKB. NFKB was originally identified as a factor that binds to the immunoglobulin kappa light chain enhancer in B cells. It was subsequently found in non-B cells in an inactive cytoplasmic form consisting of NFkappaB bound to IkappaB. NFkappaB was originally identified as a heterodimeric DNA binding protein complex consisting of p65 (RelA) and p50 (NFKB1) subunits. Other identified subunits include p52 (NFKB2), c-Rel, and RelB. The p65, cRel, and RelB subunits are responsible for transactivation. The p50 and p52 subunits possess DNA binding activity but limited ability to transactivate. p52 has been reported to form transcriptionally active heterodimers with the NFkappaB subunit p65, similar to p50/p65 heterodimers. The heterodimers of p52/p65 and p50/p65 are regulated by physical inactivation in the cytoplasm by IkB-a. IkB-a binds to the p65 subunit, preventing nuclear localization and DNA binding. Low levels of p52 and p50 homodimers can also exist in cells.

  •  実験手法
    Enzyme-linked immunosorbent assay. Read more
    Immunohistochemistry 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
    ChIP-seq (ab)
    Read more
    ChIP-qPCR (ab)
    Read more
  •  資料
    Datasheet NFKBp65 C15310256 DATASHEET
    Datasheet description
    Antibodies you can trust POSTER
    Epigenetic research tools have evolved over time from endpoint PCR to qPCR to the analyses of lar...
    Epigenetic Antibodies Brochure BROCHURE
    More than in any other immuoprecipitation assays, quality antibodies are critical tools in many e...
  •  Safety sheets
    NFKB p65 antibody SDS GB en Download
    NFKB p65 antibody SDS US en Download
    NFKB p65 antibody SDS BE nl Download
    NFKB p65 antibody SDS BE fr Download
    NFKB p65 antibody SDS FR fr Download
    NFKB p65 antibody SDS ES es Download
    NFKB p65 antibody SDS DE de Download
    NFKB p65 antibody SDS JP ja Download
  •  出版物

    How to properly cite this product in your work

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

    Using our products in your publication? Let us know!

    PBRM1-deficient PBAF complexes target aberrant genomic loci to activatethe NF-κB pathway in clear cell renal cell carcinoma.
    Yao X. et al.
    PBRM1 encodes an accessory subunit of the PBAF SWI/SNF chromatin remodeller, and the inactivation of PBRM1 is a frequent event in kidney cancer. However, the impact of PBRM1 loss on chromatin remodelling is not well examined. Here we show that, in VHL-deficient renal tumours, PBRM1 deficiency results in ectopic PBAF...

    Vitamin C enhances NF-κB-driven epigenomic reprogramming andboosts the immunogenic properties of dendritic cells.
    Morante-Palacios O. et al.
    Dendritic cells (DCs), the most potent antigen-presenting cells, are necessary for effective activation of naïve T cells. DCs' immunological properties are modulated in response to various stimuli. Active DNA demethylation is crucial for DC differentiation and function. Vitamin C, a known cofactor of ten-eleven...

    The CDK4/6-EZH2 pathway is a potential therapeutic target for psoriasis.
    Müller A, Dickmanns A, Resch C, Schäkel K, Hailfinger S, Dobbelstein M, Schulze-Osthoff K, Kramer D
    Psoriasis is a frequent inflammatory skin disease characterized by keratinocyte hyperproliferation and a disease-related infiltration of immune cells. Here, we identified a novel pro-inflammatory signaling pathway driven by the cyclin-dependent kinases (CDK) 4 and 6 and the methyltransferase EZH2 as a valid target f...

    AP-1 activity induced by co-stimulation is required for chromatin opening during T cell activation.
    Yukawa M, Jagannathan S, Vallabh S, Kartashov AV, Chen X, Weirauch MT, Barski A
    Activation of T cells is dependent on the organized and timely opening and closing of chromatin. Herein, we identify AP-1 as the transcription factor that directs most of this remodeling. Chromatin accessibility profiling showed quick opening of closed chromatin in naive T cells within 5 h of activation. These newly...

    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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