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    mkk6-antibody-ep557y-ab33866.pdf

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Cell Biology Cell Cycle Kinases/Phosphatases Other
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Validated using a knockout cell lineRabMAb

Anti-MKK6 antibody [EP557Y] (ab33866)

  • Datasheet
  • SDS
Submit a review Submit a question References (4)

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

Key features and details

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

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Secondary
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Protein
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Recombinant Human MKK6 protein (ab95252)

View more associated products

Overview

  • Product name

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

    Rabbit monoclonal [EP557Y] to MKK6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

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

    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.

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP557Y
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Other
    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • Other
    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Other
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Immunology
    • Innate Immunity
    • TLR Signaling

Associated products

  • Alternative Versions

    • Anti-MKK6 antibody [EP557Y] - BSA and Azide free (ab269878)
  • Compatible Secondaries

    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
  • Recombinant Protein

    • Recombinant Human MKK6 protein (ab95252)

Applications

The Abpromise guarantee

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.
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.
  • Target information above from: UniProt accession P52564 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5608 Human
    • Entrez Gene: 26399 Mouse
    • Entrez Gene: 114495 Rat
    • Omim: 601254 Human
    • SwissProt: P52564 Human
    • SwissProt: P70236 Mouse
    • Unigene: 463978 Human
    • Unigene: 14487 Mouse
    • 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

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (4)

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

    ab33866 has been referenced in 4 publications.

    • Norheim F  et al. Genetic regulation of liver lipids in a mouse model of insulin resistance and hepatic steatosis. Mol Syst Biol 17:e9684 (2021). PubMed: 33417276
    • Zhang X  et al. MicroRNA-375 prevents TGF-ß-dependent transdifferentiation of lung fibroblasts via the MAP2K6/P38 pathway. Mol Med Rep 22:1803-1810 (2020). PubMed: 32582987
    • 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

    Customer reviews and Q&As

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