Overview

  • Product name

    Anti-NEK7 antibody [EPR4900]
    See all NEK7 primary antibodies
  • Description

    Rabbit monoclonal [EPR4900] to NEK7
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues in Human NEK7 (UniProt ID: Q8TDX7)

  • Positive control

    • Jurkat, A549, C6, RAW264.7, PC12 and NIH/3T3 cell lysates.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133514 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
WB 1/10000. Predicted molecular weight: 35 kDa.
IP 1/10 - 1/100.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Tissue specificity

      Highly expressed in lung, muscle, testis, brain, heart, liver, leukocyte and spleen. Lower expression in ovary, prostate and kidney. No expression seen in small intestine.
    • Sequence similarities

      Belongs to the protein kinase superfamily. NEK Ser/Thr protein kinase family. NIMA subfamily.
      Contains 1 protein kinase domain.
    • Cellular localization

      Cytoplasm.
    • Information by UniProt
    • Database links

    • Alternative names

      • EC 2.7.11.1 antibody
      • NEK7 antibody
      • NEK7_HUMAN antibody
      • Never in mitosis A-related kinase 7 antibody
      • Never in mitosis gene a-related kinase 7 antibody
      • NIMA (never in mitosis gene a) related kinase 7 antibody
      • NIMA (never in mitosis gene a)-related expressed kinase 7 antibody
      • NimA related protein kinase 7 antibody
      • NIMA-related kinase 7 antibody
      • NimA-related protein kinase 7 antibody
      • Serine/threonine-protein kinase Nek7 antibody
      see all

    Images

    • ab133514 immunoprecipitating NEK7. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/20 and VeriBlot for IP Detection Reagent (HRP) (ab131366) at a dilution of 1/1000.

      Lane 1: H/3T3 (Mouse embryonic fibroblast) whole cell lysate 10ug
      Lane 2: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab133514 in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

    • All lanes : Anti-NEK7 antibody [EPR4900] (ab133514) at 1/10000 dilution

      Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
      Lane 2 : C6 (Rat glial tumor glial cell) whole cell lysate
      Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 35 kDa

    • Lane 1: Wild-type HAP1 cell lysate (40 µg)
      Lane 2: NEK7 knockout HAP1 cell lysate (40 µg)
      Lane 3: A549 cell lysate (20 µg)
      Lane 4: Jurkat cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab133514 observed at 35 kDa. Red - loading control, ab18058, observed at 124 kDa.

      ab133514 was shown to specifically react with NEK7 when NEK7 knockout samples were used. Wild-type and NEK7 knockout samples were subjected to SDS-PAGE. Ab133514 and ab18058 (loading control to Vinculin) were diluted at 1/10000 and 1/10,000 dilution respectively and incubated overnight at 4C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

    • All lanes : Anti-NEK7 antibody [EPR4900] (ab133514) at 1/10000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : A549 cell lysate
      Lane 3 : C6 cell lysate
      Lane 4 : RAW264.7 cell lysate
      Lane 5 : PC12 cell lysate
      Lane 6 : NIH/3T3 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat-anti-rabbit HRP at 1/2000 dilution

      Developed using the ECL technique.

      Predicted band size: 35 kDa
      Observed band size: 32 kDa
      why is the actual band size different from the predicted?

    References

    This product has been referenced in:

    • Hoss F  et al. Alternative splicing regulates stochastic NLRP3 activity. Nat Commun 10:3238 (2019). Read more (PubMed: 31324763) »
    • Torp MK  et al. Mammalian Target of Rapamycin (mTOR) and the Proteasome Attenuates IL-1ß Expression in Primary Mouse Cardiac Fibroblasts. Front Immunol 10:1285 (2019). Read more (PubMed: 31244838) »
    See all 11 Publications for this product

    Customer reviews and Q&As

    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (immortalised bone marrow-derived macrophage)
    Permeabilization
    Yes - 0.1% Triton X-100
    Specification
    immortalised bone marrow-derived macrophage
    Blocking step
    Fish gelatin + BSA + casein as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
    Fixative
    Paraformaldehyde

    Dr. James Harris

    Verified customer

    Submitted Jun 02 2017

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