Overview

  • Product name

    Anti-MEK3 + MEK6 antibody [EPR17345]
    See all MEK3 + MEK6 primary antibodies
  • Description

    Rabbit monoclonal [EPR17345] to MEK3 + MEK6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WB, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human MEK3 + MEK6 aa 150 to the C-terminus. The exact sequence is proprietary. Also SwissProt ID: P52564 (MEK6)
    Database link: P46734

  • Positive control

    • WB: Human MEK6 recombinant protein fragment. HepG2, HeLa, PC-12, NIH/3T3 and Jurkat whole cell lysates. Rat heart and liver lysates. Mouse heart, kidney and liver lysates. IHC-P: Human, Rat and Mouse skeletal muscle tissue. HeLa cells. HeLa whole cell extract. IP: HeLa
  • 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab181555 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
ICC/IF 1/100.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/2000. Detects a band of approximately 34, 37, 39 kDa (predicted molecular weight: 37, 39 kDa).
IP 1/140.

Target

  • Function

    Dual specificity kinase. Is activated by cytokines and environmental stress in vivo. Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in the MAP kinase p38.
  • Tissue specificity

    Abundant expression is seen in the skeletal muscle. It is also widely expressed in other tissues.
  • Involvement in disease

    Defects in MAP2K3 may be involved in colon cancer.
  • 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

    Autophosphorylated. Phosphorylation on Ser-218 and Thr-222 by MAP kinase kinase kinases regulates positively the kinase activity. Phosphorylated by TAOK2.
    Yersinia yopJ may acetylate Ser/Thr residues, preventing phosphorylation and activation, thus blocking the MAPK signaling pathway.
  • Information by UniProt
  • Database links

  • Alternative names

    • Dual specificity mitogen activated protein kinase kinase 6 antibody
    • Dual specificity mitogen-activated protein kinase kinase 3 antibody
    • MAP kinase kinase 3 antibody
    • MAP kinase kinase 6 antibody
    • map2k3 antibody
    • MAP2K6 antibody
    • MAPK/ERK kinase 3 antibody
    • MAPK/ERK kinase 6 antibody
    • MAPKK 3 antibody
    • MAPKK 6 antibody
    • MAPKK3 antibody
    • MAPKK6 antibody
    • MEK 3 antibody
    • MEK 6 antibody
    • Mitogen activated protein kinase kinase 3 antibody
    • Mitogen activated protein kinase kinase 6 antibody
    • MKK3 antibody
    • MKK6 antibody
    • MP2K3_HUMAN antibody
    • PRKMK3 antibody
    • PRKMK6 antibody
    • Protein kinase, mitogen activated, kinase 6 (MAP kinase kinase 6) antibody
    • SAPK kinase 2 antibody
    • SAPKK 3 antibody
    • SAPKK-2 antibody
    • SAPKK2 antibody
    • SAPKK3 antibody
    • Stress activated protein kinase kinase 3 antibody
    • Stress-activated protein kinase kinase 2 antibody
    see all

Images

  • All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/50000 dilution

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysates
    Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
    Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 37, 39 kDa
    Observed band size: 39,37,34 kDa
    why is the actual band size different from the predicted?



    Based on the sequence analysis, ab181555 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms and MEK6.

     

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling MEK3 + MEK6 with ab181555 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Cytoplasm and nuclear staining on HeLa cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:-
    1. ab181555 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution.

  • Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/10000 dilution + Human MEK6 recombinant protein fragment at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 37, 39 kDa
    Observed band size: 22 kDa why is the actual band size different from the predicted?



    Recombinant protein covers 139aa-334aa with a His tag (22kDa).

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/5000 dilution

    Lane 1 : Rat liver lysates
    Lane 2 : Mouse liver lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 37, 39 kDa
    Observed band size: 39,37,34 kDa why is the actual band size different from the predicted?



    Based on the sequence analysis, ab181555 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms and MEK6.

     

    Blocking/Dilution buffer: 5% NFDM/TBST.

     

  • All lanes : Anti-MEK3 + MEK6 antibody [EPR17345] (ab181555) at 1/2000 dilution

    Lane 1 : Mouse heart lysates
    Lane 2 : Mouse kidney lysates
    Lane 3 : Rat heart lysates
    Lane 4 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysates

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 37, 39 kDa
    Observed band size: 39,37,34 kDa why is the actual band size different from the predicted?



    Based on the sequence analysis, ab181555 recognizes 3 isoforms of MEK3 ranging from 36kDa to 40KDa. The multi-bands should be MEK3 isoforms and MEK6.

     

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Human skeletal muscle is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Mouse skeletal muscle is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling MEK3 + MEK6 with ab181555 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on Rat skeletal muscle is observed. Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary ab.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • MEK3 + MEK6 were immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181555 at 1/140 dilution. Western blot was performed from the immunoprecipitate using ab181555 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

  • MEK3 + MEK6 were immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab181555 at 1/140 dilution. Western blot was performed from the immunoprecipitate using ab33866 (Rabbit monoclonal [EP557Y] to MEK6) at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution. Lane 1: HeLa whole cell extract. Lane 2: PBS instead of HeLa whole cell extract.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

     

References

ab181555 has not yet been referenced specifically in any publications.

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