製品の概要

  • 製品名Anti-NF-kB p65 (phospho S536) antibody
    NF-kB p65 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to NF-kB p65 (phospho S536)
  • アプリケーション適用あり: ICC/IF, WB, IHC-Pmore details
    適用なし: IP
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Guinea pig, Dog, Pig, Chimpanzee, Rhesus monkey, Gorilla, Orangutan
  • 免疫原

    Synthetic peptide corresponding to Human NF-kB p65. around the phosphorylation site of Serine 536.

  • ポジティブ・コントロール
    • WB: Jurkat whole cell lysate treated with TNF alpha and Calyculin A. IHC_P: Breast carcinoma, Lung Adenocarcinoma, Ovarian carcinoma

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab86299 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 1/250 - 1/1000.
WB 1/2000 - 1/10000. Predicted molecular weight: 60 kDa.
IHC-P 1/500 - 1/2000.

Epitope retrieval with Tris-EDTA pH9.0 is recommended

  • 追加情報Is unsuitable for IP.
  • ターゲット情報

    • 機能NF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1.
    • 配列類似性Contains 1 RHD (Rel-like) domain.
    • ドメインthe 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
    • 翻訳後修飾Ubiquitinated, leading to its proteasomal degradation. Degradation is required for termination of NF-kappa-B response.
      Monomethylated at Lys-310 by SETD6. Monomethylation at Lys-310 is recognized by the ANK repeats of EHMT1 and promotes the formation of repressed chromatin at target genes, leading to down-regulation of NF-kappa-B transcription factor activity. Phosphorylation at Ser-311 disrupts the interaction with EHMT1 without preventing monomethylation at Lys-310 and relieves the repression of target genes.
      Phosphorylation at Ser-311 disrupts the interaction with EHMT1 and promotes transcription factor activity (By similarity). Phosphorylation on Ser-536 stimulates acetylation on Lys-310 and interaction with CBP; the phosphorylated and acetylated forms show enhanced transcriptional activity.
      Reversibly acetylated; the acetylation seems to be mediated by CBP, the deacetylation by HDAC3. Acetylation at Lys-122 enhances DNA binding and impairs association with NFKBIA. Acetylation at Lys-310 is required for full transcriptional activity in the absence of effects on DNA binding and NFKBIA association. Acetylation can also lower DNA-binding and results in nuclear export. Interaction with BRMS1 promotes deacetylation of 'Lys-310'.
    • 細胞内局在Nucleus. Cytoplasm. Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalized with RELA in the nucleus upon TNF-alpha induction.
    • Information by UniProt
    • 参照データベース
    • 別名
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      • v rel reticuloendotheliosis viral oncogene homolog A (avian) antibody
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      see all

    Anti-NF-kB p65 (phospho S536) antibody 画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat lung tissue labeling NF-kB p65 (phospho S536) with ab86299 at 1/20 dilution. Tissue sections were fixed in 6%formalin and embedded into paraffin, 5 µm thin sections were cut with a microtome. Sections were stained with haematoxylin–eosin. Slides were deparaffinized in xilene, rehydrated in graded ethanol series, and washed in distilled water. Heat induced epitope retrieval was performed by boiling the tissue sections in citrate buffer in a microwave oven at 750 W followed by cooling at room temperature for 20 minutes. Slides were washed in tris buffered saline (TBS) solution (pH = 7,6) followed by blocking of endogenous peroxidase for 10 minutes at room temperature. Slides were washed in TBS. Nonspecific sites were blocked for 10 minutes at room temperature. Without washing, the primary antibody ab86299 was applied. Incubation with the primary antibody was performed for 1 hour at room temperature followed by washing in TBS. Anti-rabbit secondary antibody was applied for 30 minutes at room temperature followed by repeated washing in TBS.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian carcinoma tissue labelling NF-kB p65 (phospho S536) with ab86299 at 1/200 (1µg/ml) and 1/1000 (0.2µg/ml). Detection: DAB.
    • All lanes : Anti-NF-kB p65 (phospho S536) antibody (ab86299) at 0.04 µg/ml

      Lane 1 : Jurkat whole cell lysate treated with TNF alpha and Calyculin A
      Lane 2 : Jurkat whole cell lysate mock treated

      Lysates/proteins at 50 µg per lane.

      Developed using the ECL technique

      Predicted band size : 60 kDa
      Observed band size : 70 kDa (why is the actual band size different from the predicted?)


      Exposure time : 30 seconds
    • IHC image of NF-kB p65 (phospho S536) staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab86299, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    Anti-NF-kB p65 (phospho S536) antibody (ab86299) 使用論文

    This product has been referenced in:
    • Kang K  et al. Carnosic acid slows photoreceptor degeneration in the Pde6b(rd10) mouse model of retinitis pigmentosa. Sci Rep 6:22632 (2016). WB . Read more (PubMed: 26961159) »
    • Koon HW  et al. Probiotic Saccharomyces boulardii CNCM I-745 prevents outbreak-associated Clostridium difficile-associated cecal inflammation in hamsters. Am J Physiol Gastrointest Liver Physiol 311:G610-G623 (2016). Read more (PubMed: 27514478) »

    See all 15 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 30 µg
    Gel Running Conditions Reduced Denaturing (10%)
    Sample Human Cell lysate - whole cell (LS174t)
    Specification LS174t
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    投稿 Jan 10 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample Mouse Tissue sections (lung tissue)
    Antigen retrieval step Heat mediated - Buffer/Enzyme Used: citrate buffer PH6.0
    Permeabilization No
    Specification lung tissue
    Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative 10% formalin
    Username

    Abcam user community

    Verified customer

    投稿 Jul 06 2016

    Application Immunohistochemistry (Frozen sections)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
    Sample Mouse Tissue sections (retina)
    Specification retina
    Permeabilization Yes - Triton x100
    Fixative Paraformaldehyde
    Username

    Dr. Minzhong Yu

    Verified customer

    投稿 Feb 27 2015

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