Overview

  • Product name
    Anti-NEK9 antibody [EP7361]
    See all NEK9 primary antibodies
  • Description
    Rabbit monoclonal [EP7361] to NEK9
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human NEK9 (N terminal). The exact sequence is proprietary.

  • Positive control
    • NIH3T3, HepG2, HeLa, and THP-1 cell lysates; Human kidney tissue; HeLa cells; Rat spleen.
  • 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 ab138488 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
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/1000 - 1/10000. Predicted molecular weight: 107 kDa.
IHC-P 1/50 - 1/100.

The mouse and rat recommendation is based on the WB results. This antibody may not be suitable for IHC with mouse or rat samples.

See IHC antigen retrieval protocols.

ICC/IF 1/100.

For unpurified use at 1/250 - 1/500.

  • Application notes
    Is unsuitable for IP.
  • Target

    • Function
      Pleiotropic regulator of mitotic progression, participating in the control of spindle dynamics and chromosome separation. Phosphorylates different histones, myelin basic protein, beta-casein, and BICD2. Phosphorylates histone H3 on serine and threonine residues and beta-casein on serine residues. Important for G1/S transition and S phase progression.
    • Tissue specificity
      Most abundant in heart, liver, kidney and testis. Also expressed in smooth muscle cells and fibroblasts.
    • Sequence similarities
      Belongs to the protein kinase superfamily. NEK Ser/Thr protein kinase family. NIMA subfamily.
      Contains 1 protein kinase domain.
      Contains 6 RCC1 repeats.
    • Developmental stage
      Expression varied mildly across the cell cycle, with highest expression observed in G1 and stationary-phase cells.
    • Domain
      Dimerizes through its coiled-coil domain.
    • Post-translational
      modifications
      Autophosphorylated on serine and threonine residues. When complexed with FACT, exhibits markedly elevated phosphorylation on Thr-210. During mitosis, not phosphorylated on Thr-210. Phosphorylated by CDK1 in vitro.
    • Cellular localization
      Cytoplasm. Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • KIAA1995 antibody
      • APUG antibody
      • DKFZp434D0935 antibody
      • LCCS10 antibody
      • MGC138306 antibody
      • MGC16714 antibody
      • NC antibody
      • NEK 9 antibody
      • Nek8 antibody
      • nek9 antibody
      • NEK9_HUMAN antibody
      • Nercc 1 kinase antibody
      • NERCC antibody
      • NERCC1 antibody
      • Nercc1 kinase antibody
      • Never in mitosis A related kinase 9 antibody
      • Never in mitosis A-related kinase 9 antibody
      • NIMA (never in mitosis gene a) related kinase 9 antibody
      • NIMA related kinase 8 antibody
      • NIMA related kinase 9 antibody
      • NIMA related kinase Nek8 antibody
      • NimA related protein kinase 9 antibody
      • NIMA-related kinase 8 antibody
      • NimA-related protein kinase 9 antibody
      • Serine/threonine protein kinase Nek9 antibody
      • Serine/threonine-protein kinase Nek9 antibody
      see all

    Images

    • Anti-NEK9 antibody [EP7361] (ab138488) at 1000 cells (purified) + Rat spleen lysates at 15 µg

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

      Predicted band size: 107 kDa
      Observed band size: 120 kDa
      why is the actual band size different from the predicted?



      Blocking and diluting buffer: 5% NFDM/TBST

    • Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling NEK9 with Purified ab138488 at 1:100 dilution (3.8μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Sonic Hedgehog with purified ab53281 at 1/20 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - cells without incubation with primary antibody and secondary antibody (Blue).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human testis tissue sections labeling NEK9 with Purified ab138488 at 1:100 dilution (3.84 μg/ml). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    • All lanes : Anti-NEK9 antibody [EP7361] (ab138488) at 1/5000 dilution (purified)

      Lane 1 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates
      Lane 2 : THP-1 (Human monocytic leukemia monocyte) whole cell lysates

      Lysates/proteins at 15 µg per lane.

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

      Predicted band size: 107 kDa
      Observed band size: 120 kDa why is the actual band size different from the predicted?



      Blocking and diluting buffer: 5% NFDM/TBST

    • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling NEK9 with unpurified ab138488 at 1/50 dilution.

    • Immunofluorescent analysis of HeLa cells labelling NEK9 with unpurified ab138488 at 1/250 dilution.

    • All lanes : Anti-NEK9 antibody [EP7361] (ab138488) at 1/1000 dilution (unpurified)

      Lane 1 : NIH3T3 cell lysate
      Lane 2 : HepG2 cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : THP1 cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 107 kDa

    References

    This product has been referenced in:
    See all 5 Publications for this product

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