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Anti-NFkB Inducing Kinase NIK 抗体 (ab19144)

Submit a review Q&A (2)References (4)
Western blot - Anti-NFkB Inducing Kinase NIK antibody (ab19144)
  • Immunocytochemistry/ Immunofluorescence - Anti-NFkB Inducing Kinase NIK antibody (ab19144)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB Inducing Kinase NIK antibody (ab19144)

Key features and details

  • Rabbit polyclonal to NFkB Inducing Kinase NIK
  • Suitable for: IHC-P, ICC/IF, WB
  • Reacts with: Human
  • Isotype: IgG

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-NFkB Inducing Kinase NIK antibody [3H31L48] (ab203568)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 製品名

    Anti-NFkB Inducing Kinase NIK antibody
    NFkB Inducing Kinase NIK 一次抗体 製品一覧

  • 製品の詳細

    Rabbit polyclonal to NFkB Inducing Kinase NIK

  • 由来種

    Rabbit

  • 特異性

    This antibody recognises human NFkB Inducing Kinase NIK.

  • アプリケーション

    適用あり: IHC-P, ICC/IF, WBmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide:

    SFAWSWRVKHGQLENRP

    , corresponding to amino acids 931-947 of Human NFkB Inducing Kinase NIK
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI

  • ポジティブ･コントロール

    • Whole cell lysate of 293.

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab19144の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 1 - 5 µg/ml.
WB
1/500. Detects a band of approximately 105 kDa (predicted molecular weight: 105.4 kDa).
特記事項
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 1 - 5 µg/ml.
WB
1/500. Detects a band of approximately 105 kDa (predicted molecular weight: 105.4 kDa).

ターゲット情報

  • 機能

    Lymphotoxin beta-activated kinase which seems to be exclusively involved in the activation of NF-kappa-B and its transcriptional activity. Promotes proteolytic processing of NFKB2/P100, which leads to activation of NF-kappa-B via the non-canonical pathway. Could act in a receptor-selective manner.

  • 組織特異性

    Weakly expressed in testis, small intestine, spleen, thymus, peripheral blood leukocytes, prostate, ovary and colon.

  • 配列類似性

    Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
    Contains 1 protein kinase domain.

  • 翻訳後修飾

    Autophosphorylated. Phosphorylation at Thr-559 is required to activates its kinase activity and 'Lys-63'-linked polyubiquitination. Phosphorylated by CHUK/IKKA leading to MAP3K14 destabilization.
    Ubiquitinated. Undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. 'Lys-48'-linked polyubiquitination leads to its degradation by the proteasome, while 'Lys-63'-linked polyubiquitination stabilizes and activates it.

  • 細胞内局在

    Cytoplasm.
  • Information by UniProt

  • 参照データベース

    • 別名

      • MAP3K14 antibody
      • FTDCR1B antibody
      • HS antibody
      • HsNIK antibody
      • M3K14_HUMAN antibody
      • Map3k14 antibody
      • Mitogen activated protein kinase kinase kinase 14 antibody
      • Mitogen-activated protein kinase kinase kinase 14 antibody
      • NF kappa beta inducing kinase antibody
      • NF-kappa-beta-inducing kinase antibody
      • NIK antibody
      • Serine/threonine-protein kinase NIK antibody
      see all

    画像

    • Western blot - Anti-NFkB Inducing Kinase NIK antibody (ab19144)
      Western blot - Anti-NFkB Inducing Kinase NIK antibody (ab19144)
      Anti-NFkB Inducing Kinase NIK antibody (ab19144) at 1/500 dilution + NIK-transfected 293 cell lysate

      Predicted band size: 105.4 kDa

    • Immunocytochemistry/ Immunofluorescence - Anti-NFkB Inducing Kinase NIK antibody (ab19144)
      Immunocytochemistry/ Immunofluorescence - Anti-NFkB Inducing Kinase NIK antibody (ab19144)
      ICC/IF image of ab19144 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab19144, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB Inducing Kinase NIK antibody (ab19144)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB Inducing Kinase NIK antibody (ab19144)
      IHC image of ab19144 staining in human cervix 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 ab19144, 5µg/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.

    参考文献 (4)

    ab19144 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab19144 は 4 報の論文で使用されています。

    • Li J  et al. Hypoxic colorectal cancer-derived extracellular vesicles deliver microRNA-361-3p to facilitate cell proliferation by targeting TRAF3 via the noncanonical NF-?B pathways. Clin Transl Med 11:e349 (2021). PubMed: 33784010
    • Das R  et al. Lymphotoxin-ß receptor-NIK signaling induces alternative RELB/NF-?B2 activation to promote metastatic gene expression and cell migration in head and neck cancer. Mol Carcinog 58:411-425 (2019). PubMed: 30488488
    • Maracle CX  et al. Noncanonical NF-?B signaling in microvessels of atherosclerotic lesions is associated with inflammation, atheromatous plaque morphology and myocardial infarction. Atherosclerosis 270:33-41 (2018). IHC-P ; Human . PubMed: 29407886
    • Maracle CX  et al. Silencing NIK potentiates anti-VEGF therapy in a novel 3D model of colorectal cancer angiogenesis. Oncotarget 9:28445-28455 (2018). PubMed: 29983872

    レビューと Q&A

    Show All レビュー Q&A
    レビューを送る 質問を送る

    1-2 of 2 Abreviews or Q&A

    Question

    Do non-transfected 293 cells express the NIK protein? What would be a good positive control for NIK? What is the best NIK antibody?

    Read More
    Answer

    Thank you very much for your call earlier this week and for your patience while I've been in touch with my colleagues regarding your enquiry.

    I've confirmed that ab37712 was tested with non-transfected 293 lysate, however ab19144 was actuallytested with transfected 293 lysate and the datasheet is currently being updated to reflect this. The lab has one publication on filein whichfull-length NIK cDNA wasfound in a293 cell cDNA library, so it does seem like this cell line endogenously expresses NIK-

    http://www.pnas.org/content/94/18/9792.full

    It's possible that certain antibodies and/or protocols will detect transfected NIK more efficiently or that the endogenous titer is low enough not to be detected by some antibodies. As a positive control, it may be better to use 293 lysate with one of the antibodies that has been tested with non-transfected lysate (ab37712, ab78273, ab8111) but if you're using one of the other NIK antibodies, I'd suggest using a recombinant NIK protein as a positive control.

    I recommend either ab7204 or ab8111, as these have each beenreferenced in a publication-

    https://www.abcam.com/NFkB-Inducing-Kinase-NIK-antibody-ab7204.html

    https://www.abcam.com/NFkB-Inducing-Kinase-NIK-antibody-ab8111.html

    We do have full-length recombinant NIK that can be used as a positive control with either of these antibodies-

    https://www.abcam.com/NIK-protein-Tagged-ab132206.html

    Please note that we do guarantee all of these antibodies in Western blot with human samples, so if you do have any trouble with the antibody that you choose, we'd be happy to refund or replace it for you. Also, we are running a promotion until the end of September in which if you're purchasing 3 items on one order, you can receive a fourth item for free.

    I hope that this information will be useful, but if you have further questions or need anything else, please let me know and I'll be happy to help.

    Read More

    Question

    thank you for your quick answer. I actually attached the foto (powerpoint file) of a western blot analysis using the NIK ab19144 to the original enquiry. This time an TIFF file is attached to document my results.

    Read More
    Answer

    I have received feedback from the laboratory who understand why you are concerned about your results as the positive control normally gives a very nice strong band at 105kDa. I have added the image of the lab on the datasheet to show you our results. We also see lower bands which we believe are breakdown products of NIK and the only possible explanation for your problem is that in your samples the protein is not protected enough against breakdown by proteases. Could you check that the concentration of inhibitors is high enough and that the inhibitors have not gone off? I was unable to find a recent order for Germany for this antibody (I wanted to check for possible shipping delays which would suggest a damaged vial), did you purchase this antibody a long time ago? If so it may be that it has been damaged during the storage unfortunately. I hope this information will help you, please do not hesitate to contact me if I can be of further help,

    Read More

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