Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Key features and details
- Rabbit polyclonal to MEK1 + MEK2 (phospho S222)
- Suitable for: ICC/IF, WB
- Reacts with: Mouse, Human
- Isotype: IgG
Overview
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Product name
Anti-MEK1 + MEK2 (phospho S222) antibody
See all MEK1 + MEK2 primary antibodies -
Description
Rabbit polyclonal to MEK1 + MEK2 (phospho S222) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Chicken, Xenopus laevis, Chimpanzee -
Immunogen
Synthetic peptide corresponding to Human MEK1 + MEK2 (phospho S222).
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Positive control
- ICC/IF: Mouse embryonic fibroblasts WB: NIH3T3 cells
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
BSA is IgG and protease free -
Concentration information loading...
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Purity
Immunogen affinity purified -
Purification notes
Purified from rabbit serum by sequential epitope-specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated MEK 1 + 2. The final product is generated by affinity chromatography using a MEK 1 + 2 derived peptide that is phosphorylated at serine 222. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab4750 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 |
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ICC/IF |
Use at an assay dependent concentration.
Assay dependent. |
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WB |
1/1000. Predicted molecular weight: 43.3 kDa.
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Notes |
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ICC/IF
Use at an assay dependent concentration. Assay dependent. |
WB
1/1000. Predicted molecular weight: 43.3 kDa. |
Target
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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
modificationsPhosphorylation 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
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Database links
- Entrez Gene: 5604 Human
- Entrez Gene: 5605 Human
- Entrez Gene: 26395 Mouse
- Entrez Gene: 26396 Mouse
- Entrez Gene: 379991 Xenopus laevis
- Omim: 176872 Human
- Omim: 601263 Human
- SwissProt: Q90891 Chicken
see all -
Alternative names
- AA589381 antibody
- CFC3 antibody
- Dual specificity mitogen-activated protein kinase kinase 1 antibody
see all
Images
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All lanes : Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Lane 1 : A549 whole cell extract lysate
Lane 2 : A549 whole cell lysate treated with EGF (200 ng/mL for 10 min)
Lane 3 : A549 whole cell lysate treated with Afatinib (0.5 uM for 16 h) followed by EGF (200 ng/mL for 10 min)
Lysates/proteins at 30 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 0.25 µg/ml
Predicted band size: 43.3 kDaWestern blot analysis was performed on whole cell extracts of A549 in different conditions, observed a 45 kDa band corresponding to Phospho-MEK1/MEK2 (Ser222) was observed in untreated A549 lysate, the signal increased upon EGF treatment, and decreased upon pretreatment with the EGFR antagonist, Afatinib. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard. Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.
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Lane 1 : Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Lane 1 : Untreated NIH3T3 cells
Lanes 2-5 :Anti-MEK1 + MEK2 (phospho S222) antibody (ab4750)
Secondary
Lane 1 : A goat F (ab)2 anti-rabbit IgG alkaline phosphatase
Predicted band size: 43.3 kDaPeptide Competition and Stimulation Extracts of NIH3T3 cells untreated (lane 1) or treated with 50 ng/mL PDGF for 15 minutes (2-5) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 4% BSA-TBST buffer overnight at 4°C, then incubated with the MEK1&2 (pS222) antibody for two hours at room temperature in a 1% BSA-TBST buffer, following various prior incubation conditions. After washing, the membrane was incubated with goat F (ab)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Pierce SuperSignal method. The data show that only the phosphopeptide corresponding to MEK1&2 (pS222) block the antibody signal, demonstrating the specificity of the antibody. The data also show the induction of MEK1&2 (pS222) phosphorylation by the addition of PDGF to this cell system. -
Immunofluorescent analysis of mouse embryonic fibroblasts labelled for MEK1/2 (pS222) using ab4750, phosphospecific antibody. The cell is actively dividing. Blue represents chromosomes in anaphase of mitotic cell division. Green shows mitotic spindle expressing phosphorylated MEK1/2.
Protocols
Datasheets and documents
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Datasheet download
References (1)
ab4750 has been referenced in 1 publication.
- Qu P et al. Effects of Rolling-Sliding Mechanical Stimulation on Cartilage Preserved In Vitro. Cell Mol Bioeng 12:301-310 (2019). PubMed: 31719916