Overview

  • Product name
    Anti-MEK1 antibody [Y77]
    See all MEK1 primary antibodies
  • Description
    Rabbit monoclonal [Y77] to MEK1
  • Host species
    Rabbit
  • Specificity
    The antibody does not crossreact with other MAP kinase kinase family members.
  • Tested applications
    Suitable for: ICC/IF, WB, Flow Cyt, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human MEK1 aa 350-450 (C terminal). The exact sequence is proprietary.

  • Positive control
    • WB: Wild-type HAP1 whole cell lysate; A431, Jurkat, HeLa and HepG2 whole cell lysates. IHC-P: Urinary bladder carcinoma tissue, human kidney tissue. ICC/IF: HeLa, A431 ells. IP: Jurkat cell lysate FC: HeLa cells
  • General notes

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

     

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32576 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 1/50.
WB 1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 43 kDa).
Flow Cyt 1/30 - 1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/20 - 1/50.
IHC-P 1/100.

See IHC antigen retrieval protocols.

Target

  • Function
    Catalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in MAP kinases. Activates ERK1 and ERK2 MAP kinases.
  • Tissue specificity
    Widely expressed, with extremely low levels in brain.
  • Involvement in disease
    Defects in MAP2K1 are a cause of cardiofaciocutaneous syndrome (CFC syndrome) [MIM:115150]; also known as cardio-facio-cutaneous syndrome. CFC syndrome is characterized by a distinctive facial appearance, heart defects and mental retardation. Heart defects include pulmonic stenosis, atrial septal defects and hypertrophic cardiomyopathy. Some affected individuals present with ectodermal abnormalities such as sparse, friable hair, hyperkeratotic skin lesions and a generalized ichthyosis-like condition. Typical facial features are similar to Noonan syndrome. They include high forehead with bitemporal constriction, hypoplastic supraorbital ridges, downslanting palpebral fissures, a depressed nasal bridge, and posteriorly angulated ears with prominent helices. The inheritance of CFC syndrome is autosomal dominant.
  • 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
    Phosphorylation on Ser/Thr by MAP kinase kinase kinases (RAF or MEKK1) regulates positively the kinase activity.
    Acetylation by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway.
  • Information by UniProt
  • Database links
  • Alternative names
    • Dual specificity mitogen activated protein kinase kinase 1 antibody
    • Dual specificity mitogen-activated protein kinase kinase 1 antibody
    • ERK activator kinase 1 antibody
    • MAP kinase kinase 1 antibody
    • MAP kinase/Erk kinase 1 antibody
    • MAP2K1 antibody
    • MAPK/ERK kinase 1 antibody
    • MAPKK 1 antibody
    • MAPKK1 antibody
    • MEK 1 antibody
    • Mek1 antibody
    • MEKK1 antibody
    • Mitogen activated protein kinase kinase 1 antibody
    • MKK 1 antibody
    • MKK1 antibody
    • MP2K1_HUMAN antibody
    • PRKMK1 antibody
    • Protein kinase mitogen activated kinase 1 (MAP kinase kinase 1) antibody
    • Protein kinase mitogen activated, kinase 1 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: MEK1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: A431 whole cell lysate (20 µg)
    Lane 4: HepG2 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32576 (unpurified) observed at 43 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab32576 was shown to specifically react with MEK1 in wild-type HAP1 cells as signal was lost in MEK1 knockout cells. Wild-type and MEK1 knockout samples were subjected to SDS-PAGE. Ab32576 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at  1/10000 dilution and 1/20000 dilution respectively. Blots were developed 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 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human kidney tissue sections labeling MEK1 with purified ab32576 at 1/100 dilution (3.28 µg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

  • All lanes : Anti-MEK1 antibody [Y77] (ab32576) at 1/10000 dilution (Purified)

    Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
    Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?

  • Immunocytochemistry/ Immunofluorescence analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling MEK1 with purified ab32576 at 1/50 dilution (6.6 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • ab32576 (purified) at 1/20 dilution (2ug) immunoprecipitating MEK1 in Jurkat whole cell lysate. Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 10ug
    Lane 2 (+): ab32576 & Jurkat whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32576 in Jurkat whole cell lysate
    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling MEK1 with purified ab32576 at 1/30 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling MEK1 with purified ab32576 at 1/500 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • ab32576 (unpurified) at a 1/250 dilution staining MEK1 in human urinary bladder carcinoma tissue by IHC-P.

  • Anti-MEK1 antibody [Y77] (ab32576) at 1/10000 dilution (unpurified) + Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate

    Predicted band size: 43 kDa
    Observed band size: 45 kDa why is the actual band size different from the predicted?

References

This product has been referenced in:
  • Sui H  et al. Lidocaine inhibits growth, migration and invasion of gastric carcinoma cells by up-regulation of miR-145. BMC Cancer 19:233 (2019). Read more (PubMed: 30876463) »
  • Milewska M  et al. Development of a personalized therapeutic strategy for ERBB-gene-mutated cancers. Ther Adv Med Oncol 10:1758834017746040 (2018). Read more (PubMed: 29383036) »
See all 9 Publications for this product

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