Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-MEK1 antibody [Y77] - BSA and Azide free (ab167151)

Overview

  • Product name
    Anti-MEK1 antibody [Y77] - BSA and Azide free
    See all MEK1 primary antibodies
  • Description
    Rabbit monoclonal [Y77] to MEK1 - BSA and Azide free
  • Host species
    Rabbit
  • Specificity

    The antibody does not crossreact with other MAP kinase kinase family members.

  • Tested applications
    Suitable for: WB, IP, Flow Cyt, ICC/IF, 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 and HepG2 whole cell lysates. IHC-P: Urinary bladder carcinoma tissue. ICC/IF: HeLa cells.
  • General notes

    ab167151 is a PBS-only buffer format of ab32576. Please refer to ab32576 for recommended dilutions, protocols, and image data.

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    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 ab167151 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 Use at an assay dependent concentration. Detects a band of approximately 45 kDa (predicted molecular weight: 43 kDa).
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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

ICC/IF Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32576).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32576).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32576).

  • 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).
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32576).

  • 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.
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32576).

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32576).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32576).

References

ab167151 has not yet been referenced specifically in any publications.

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