Overview

  • Product name
    Anti-NFkB Inducing Kinase NIK antibody
    See all NFkB Inducing Kinase NIK primary antibodies
  • Description
    Rabbit polyclonal to NFkB Inducing Kinase NIK
  • Host species
    Rabbit
  • Specificity
    This antibody recognises human NFkB Inducing Kinase NIK.
  • Tested applications
    Suitable for: IHC-P, ICC/IF, WBmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide:

    SFAWSWRVKHGQLENRP

    , corresponding to amino acids 931-947 of Human NFkB Inducing Kinase NIK

  • Positive control
    • Whole cell lysate of 293.

Properties

Applications

Our Abpromise guarantee covers the use of ab19144 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
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).Can be blocked with NFkB Inducing Kinase NIK peptide (ab8385).

Target

  • Function
    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.
  • Tissue specificity
    Weakly expressed in testis, small intestine, spleen, thymus, peripheral blood leukocytes, prostate, ovary and colon.
  • Sequence similarities
    Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    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.
  • Cellular localization
    Cytoplasm.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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

Images

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

    Predicted band size: 105.4 kDa

  • 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.

  • 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.

References

This product has been referenced in:
  • 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 . Read more (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). Read more (PubMed: 29983872) »
See all 2 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

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.

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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,

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