Overview

  • Product name

    Anti-MEK3 + MEK6 antibody [EPR17340]
    See all MEK3 + MEK6 primary antibodies
  • Description

    Rabbit monoclonal [EPR17340] to MEK3 + MEK6
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human MEK3 + MEK6 aa 300 to the C-terminus.
    Database link: P46734

  • Positive control

    • WB: Human MEK6 recombinant protein fragment; HeLa, HepG2, Jurkat, MCF7, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Human fetal liver lysate; Mouse and rat spleen lysates. IHC-P: Human cervix carcinoma and spleen tissues; Mouse liver tissue. ICC/IF: HeLa and HepG2 cells. FC & IP: HeLa whole cell lysate.
  • 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 ab200831 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
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 39, 37, 34 kDa (predicted molecular weight: 39, 37 kDa).
ICC/IF 1/1000.
IP 1/100.
Flow Cyt 1/150.

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 [EPR17340] (ab200831) at 1/1000 dilution

    Lane 1 : Mouse spleen lysate at 10 µg
    Lane 2 : Rat spleen lysate at 10 µg
    Lane 3 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 4 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 6 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

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

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


    Exposure time: 30 seconds


    Blocking/Dilution Buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling MEK3 + MEK6 with ab200831 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on Mouse liver tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

  • Anti-MEK3 + MEK6 antibody [EPR17340] (ab200831) at 1/10000 dilution + Human MEK6 recombinant protein fragment at 10 µg

    Predicted band size: 39, 37 kDa


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

     

    Recombinant fragment of Human MEK6 protein contains aa139-334 with His-Tag®(22kDa).

  • Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling MEK3 + MEK6 with ab200831 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

  • All lanes : Anti-MEK3 + MEK6 antibody [EPR17340] (ab200831) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate
    Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

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

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


    Exposure time: 15 seconds


    Blocking/Dilution Buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling MEK3 + MEK6 with ab200831 at 1/250 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear and cytoplasmic staining on Human spleen tissue is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

  • Anti-MEK3 + MEK6 antibody [EPR17340] (ab200831) at 1/10000 dilution + Human fetal liver lysate at 10 µg

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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


    Exposure time: 1 minute


    Blocking/Dilution Buffer: 5% NFDM/TBST.

  • Flow cytometry analysis of HeLa cells labelling MEK3 + MEK6 (red) with purified ab200831 at dilution of 1/150. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Isotype control antibody was Rabbit monoclonal IgG (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • MEK3 + MEK6 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab200831 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab200831 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 lysate 10ug (Input). Lane 2: ab200831 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200831 in Jurkat whole cell lysate.
    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 30 seconds.

References

This product has been referenced in:

  • Guo Y  et al. Downregulation of HS6ST2 by miR-23b-3p enhances matrix degradation through p38 MAPK pathway in osteoarthritis. Cell Death Dis 9:699 (2018). Read more (PubMed: 29899528) »
See 1 Publication for this product

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