Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP7361] to NEK9 - BSA and Azide free
  • Suitable for: ICC, IHC-P, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-NEK9 antibody [EP7361] - BSA and Azide free
    See all NEK9 primary antibodies

  • Description

    Rabbit monoclonal [EP7361] to NEK9 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    IHC-P
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431, HepG2, MCF7, NIH/3T3 and THP-1 cell lysates. IHC-P: Human testis and kidney tissues. ICC: HeLa cells.

  • General notes

    Ab248844 is the carrier-free version of ab138488. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab248844 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Western blot - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    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?



    This data was developed using ab138488, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Immunocytochemistry - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Immunocytochemistry - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)

    This data was developed using ab138488, the same antibody clone in a different buffer formulation.

    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 - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Flow Cytometry - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)

    This data was developed using ab138488, the same antibody clone in a different buffer formulation.

    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) - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    This data was developed using ab138488, the same antibody clone in a different buffer formulation.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 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.
  • Western blot - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Western blot - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    All lanes : Anti-NEK9 antibody [EP7361] (ab138488) at 1/1000 dilution

    Lane 1 : Wild-type A431 cell lysate
    Lane 2 : NEK9 knockout A431 cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : MCF7 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

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



    This data was developed using the same antibody clone in a different buffer formulation (ab138488).

    Lanes 1 - 4: Merged signal (red and green). Green - ab138488 observed at 120 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab138488 was shown to react with NEK9 in A431 wild-type cells in western blot. Loss of signal was observed when NEK9 knockout sample was used. Wild-type and NEK9 knockout A431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab138488 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    This data was developed using ab138488, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human kidney tissue labelling NEK9 with unpurified ab138488 at 1/50 dilution.
  • Immunocytochemistry - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Immunocytochemistry - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    This data was developed using ab138488, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of HeLa cells labelling NEK9 with unpurified ab138488 at 1/250 dilution.
  • Western blot - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Western blot - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    All lanes : Anti-NEK9 antibody [EP7361] (ab138488) at 1/5000 dilution (purified)

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

    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?



    This data was developed using ab138488, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Western blot - Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    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



    This data was developed using ab138488, the same antibody clone in a different buffer formulation.

     

  • Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)
    Anti-NEK9 antibody [EP7361] - BSA and Azide free (ab248844)

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