Overview

  • Product name

    Anti-MEK1 + MEK2 antibody [EPR16667] - BSA and Azide free
    See all MEK1 + MEK2 primary antibodies
  • Description

    Rabbit monoclonal [EPR16667] to MEK1 + MEK2 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human MEK1 + MEK2 aa 1 to the C-terminus. The exact sequence is proprietary. Also SwissProt ID: P36507
    Database link: Q02750

  • Positive control

    • WB: HeLa, 293T, A549 and A431 whole cell lysates; Human fetal brain, heart, kidney and spleen lysates; Mouse and Rat brain and heart lysates. IHC-P: Human renal medulla, Mouse lung and Rat lung tissues. ICC/IF: NIH/3T3 cells. IP: HeLa whole cell extract.
  • General notes

    Ab215263 is the carrier-free version of ab178876. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab215263 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at 4°C (up to 6 months). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Constituent: PBS
  • Concentration information loading...
  • Purity

    Affinity purified
  • Clonality

    Monoclonal
  • Clone number

    EPR16667
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab215263 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 Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Detects a band of approximately 44 kDa (predicted molecular weight: 43, 44 kDa).

Target

  • Function

    Dual specificity protein kinase which acts as an essential component of the MAP kinase signal transduction pathway. Binding of extracellular ligands such as growth factors, cytokines and hormones to their cell-surface receptors activates RAS and this initiates RAF1 activation. RAF1 then further activates the dual-specificity protein kinases MAP2K1/MEK1 and MAP2K2/MEK2. Both MAP2K1/MEK1 and MAP2K2/MEK2 function specifically in the MAPK/ERK cascade, and catalyze the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in the extracellular signal-regulated kinases MAPK3/ERK1 and MAPK1/ERK2, leading to their activation and further transduction of the signal within the MAPK/ERK cascade. Depending on the cellular context, this pathway mediates diverse biological functions such as cell growth, adhesion, survival and differentiation, predominantly through the regulation of transcription, metabolism and cytoskeletal rearrangements. One target of the MAPK/ERK cascade is peroxisome proliferator-activated receptor gamma (PPARG), a nuclear receptor that promotes differentiation and apoptosis. MAP2K1/MEK1 has been shown to export PPARG from the nucleus. The MAPK/ERK cascade is also involved in the regulation of endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC), as well as in the fragmentation of the Golgi apparatus during mitosis.
  • Tissue specificity

    Widely expressed, with extremely low levels in brain.
  • Involvement in disease

    Cardiofaciocutaneous syndrome 3
  • Sequence similarities

    Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily.
    Contains 1 protein kinase domain.
  • Domain

    The proline-rich region localized between residues 270 and 307 is important for binding to RAF1 and activation of MAP2K1/MEK1.
  • Post-translational
    modifications

    Phosphorylation at Ser-218 and Ser-222 by MAP kinase kinase kinases (RAF or MEKK1) positively regulates kinase activity. Also phosphorylated at Thr-292 by MAPK1/ERK2 and at Ser-298 by PAK. MAPK1/ERK2 phosphorylation of Thr-292 occurs in response to cellular adhesion and leads to inhibition of Ser-298 phosphorylation by PAK.
    Acetylation by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway.
  • Cellular localization

    Cytoplasm, cytoskeleton, microtubule organizing center, centrosome. Cytoplasm, cytoskeleton, microtubule organizing center, spindle pole body. Cytoplasm. Nucleus. Localizes at centrosomes during prometaphase, midzone during anaphase and midbody during telophase/cytokinesis.
  • Information by UniProt
  • Database links

  • Alternative names

    • AA589381 antibody
    • CFC3 antibody
    • Dual specificity mitogen-activated protein kinase kinase 1 antibody
    • Dual specificity mitogen-activated protein kinase kinase 2 antibody
    • EC 2.7.12.2 antibody
    • ERK activator kinase 1 antibody
    • ERK activator kinase 2 antibody
    • FLJ26075 antibody
    • MAP kinase kinase 1 antibody
    • MAP kinase kinase 2 antibody
    • MAP2K1 antibody
    • MAP2K2 antibody
    • MAPK/ERK kinase 1 antibody
    • MAPK/ERK kinase 2 antibody
    • MAPKK 1 antibody
    • MAPKK1 antibody
    • MAPKK2 antibody
    • MEK 1 antibody
    • MEK1 antibody
    • MEKK1 antibody
    • Mitogen activated protein kinase kinase 1 antibody
    • Mitogen activated protein kinase kinase 2 antibody
    • Mitogen-activated protein kinase kinase 2, p45 antibody
    • MK2 antibody
    • MKK 1 antibody
    • MKK 2 antibody
    • MKK1 antibody
    • MKK2 antibody
    • MP2K1_HUMAN antibody
    • PRKMK 1 antibody
    • PRKMK 2 antibody
    • Prkmk1 antibody
    • Prkmk2 antibody
    • protein kinase, mitogen-activated, kinase 1 (MAP kinase kinase 1) antibody
    • Protein kinase, mitogen-activated, kinase 1 antibody
    • Protein kinase, mitogen-activated, kinase 2 antibody
    see all

Images

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling MEK1 + MEK2 with ab178876 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/200 dilution (green). Cytoplasmic staining on NIH/3T3 cell line is observed. The nuclear counter stain 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. ab178876 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 1/200 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178876).

  • ab178876 staining MEK1 + MEK2 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permabilised with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled 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 (ab178876).

  • Immunohistochemical analysis of paraffin-embedded Human renal medulla tissue labeling MEK1 + MEK2 with ab178876 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Renal tubule epithelial cells show strong cytoplasmic staining with some additional nuclear staining. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178876).

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

  • Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling MEK1 + MEK2 with ab178876 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Alveolar epithelial cells show strong cytoplasmic staining with some additional nuclear staining. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178876).

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

  • Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling MEK1 + MEK2 with ab178876 at 1/500 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Alveolar epithelial cells show strong cytoplasmic staining with some additional nuclear staining. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178876).

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

  • MEK1 + MEK2 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell extract with ab178876 at 1/120 dilution. Western blot was performed from the immunoprecipitate using ab178876 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab178876).

References

This product has been referenced in:

  • Jia S  et al. Overexpression of indoleamine 2, 3-dioxygenase contributes to the repair of human airway epithelial cells inhibited by dexamethasone via affecting the MAPK/ERK signaling pathway. Exp Ther Med 16:282-290 (2018). Read more (PubMed: 29896251) »
See 1 Publication for this product

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