Validated using a knockout cell lineRabMAb

Anti-MKK6 antibody [EP557Y] (ab33866)

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Key features and details

  • Rabbit monoclonal [EP557Y] to MKK6
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

Overview

  • Product name

    Anti-MKK6 antibody [EP557Y]
    See all MKK6 primary antibodies

  • Description

    Rabbit monoclonal [EP557Y] to MKK6

  • Host species

    Rabbit

  • Specificity

    ab33866 recognises MKK6.

  • Tested applications

    Suitable for: WB, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human MKK6 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HepG2, C2C12, HeLa, HAP1, and C6 lysates IP: HepG2 cells

  • General notes

    This product was previously labelled as MEK6.

     

    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.

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab33866 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 1/1000 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 37 kDa).
IP 1/20 - 1/50.

Target

  • Function

    Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in MAP kinase p38 exclusively.

  • Tissue specificity

    Isoform 2 is only expressed in skeletal muscle. Isoform 1 is expressed in skeletal muscle, heart, and in lesser extent in liver or pancreas.

  • Sequence similarities

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

  • Post-translational
    modifications

    Weakly autophosphorylated.
    Acetylation of Ser-207 and Thr-211 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway.
  • Information by UniProt

  • Database links

    • Form

      It localizes to nucleus, cytoplasm and cytoskeleton. Note: Binds to microtubules.

    • Alternative names

      • Dual specificity mitogen activated protein kinase kinase 6 antibody
      • Dual specificity mitogen-activated protein kinase kinase 6 antibody
      • MAP kinase kinase 6 antibody
      • MAP2K6 antibody
      • MAPK/ERK kinase 6 antibody
      • MAPKK 6 antibody
      • MAPKK6 antibody
      • MEK 6 antibody
      • MEK6 antibody
      • Mitogen Activated Protein Kinase Kinase 6 antibody
      • MKK 6 antibody
      • MKK6 antibody
      • MP2K6_HUMAN antibody
      • PRKMK6 antibody
      • protein kinase antibody
      • Protein kinase mitogen activated kinase 6 antibody
      • protein kinase, mitogen-activated, kinase 6 (MAP kinase kinase 6) antibody
      • SAPK kinase 3 antibody
      • SAPKK-3 antibody
      • SAPKK3 antibody
      • SKK3 antibody
      • Stress-activated protein kinase kinase 3 antibody
      see all

    Images

    • Western blot - Anti-MKK6 antibody [EP557Y] (ab33866)
      Western blot - Anti-MKK6 antibody [EP557Y] (ab33866)

      Lanes 1, 5 and 9: Wild-type HAP1 cell lysate (20 µg)
      Lanes 2, 6 and 10: MEK6 knockout HAP1 cell lysate (20 µg)
      Lanes 3, 7 and 11: HeLa cell lysate (20 µg)
      Lanes 4, 8 and 12: A431 cell lysate (20 µg)
      Lanes 1, 2, 3 and 4: Green signal from target – ab33866 observed at 37 kDa
      Lanes 5, 6, 7 and 8: Red signal from loading control – ab18058 observed at 130 kDa
      Lanes 9, 10, 11 and 12: Merged (red and green) signal

      ab33866 (unpurified) was shown to specifically react with MEK6 when MEK6 knockout samples were used. Wild-type and MEK6 knockout samples were subjected to SDS-PAGE. ab33866 and ab18058 (loading control to Vinculin) were diluted 1/10 000 and 1/2000 respectively and incubated overnight at 4ºC. 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 h at room temperature before imaging.

    • Western blot - Anti-MKK6 antibody [EP557Y] (ab33866)
      Western blot - Anti-MKK6 antibody [EP557Y] (ab33866)
      All lanes : Anti-MKK6 antibody [EP557Y] (ab33866) at 1/1000 dilution

      Lane 1 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates
      Lane 2 : C2C12 (Mouse myoblasts myoblast) whole cell lysates
      Lane 3 : C6 (Rat glial tumor glial cell) 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: 37 kDa
      Observed band size: 36 kDa
      why is the actual band size different from the predicted?

    • Immunoprecipitation - Anti-MKK6 antibody [EP557Y] (ab33866)
      Immunoprecipitation - Anti-MKK6 antibody [EP557Y] (ab33866)
      ab33866 (purified) at 1/20 dilution (1 µg) immunoprecipitating MKK6 in HepG2 whole cell lysate.
      Lane 1 (input): HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10 µg
      Lane 2 (+): ab33866 & HepG2 whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab33866 in HepG2 whole cell lysate
      For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
      Blocking and diluting buffer: 5% NFDM/TBST.

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (2)

    Publishing research using ab33866? Please let us know so that we can cite the reference in this datasheet.

    ab33866 has been referenced in 2 publications.

    • Grun D  et al. NRP-1 interacts with GIPC1 and SYX to activate p38 MAPK signaling and cancer stem cell survival. Mol Carcinog N/A:N/A (2018). PubMed: 30456845
    • Sikorski K  et al. A high-throughput pipeline for validation of antibodies. Nat Methods 15:909-912 (2018). PubMed: 30377371

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