Recombinant Anti-MEK1 antibody [E342] (ab32091)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E342] to MEK1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-MEK1 antibody [E342]
See all MEK1 primary antibodies -
Description
Rabbit monoclonal [E342] to MEK1 -
Host species
Rabbit -
Specificity
This antibody recognises MEK1, but does not cross react with other MAP kinase kinase family members. -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Cow, Dog -
Immunogen
Synthetic peptide within Human MEK1 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: Q02750 -
Positive control
- WB: A431 cells and cell lysate. IHC-P: Human cervical carcinoma. ICC/IF: HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E342 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32091 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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Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (4) |
1/1000 - 1/5000. Detects a band of approximately 45 kDa (predicted molecular weight: 43 kDa).
|
IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/100 - 1/500.
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Notes |
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Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/5000. Detects a band of approximately 45 kDa (predicted molecular weight: 43 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/100 - 1/500. |
Target
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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
modificationsPhosphorylation 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
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Database links
- Entrez Gene: 533199 Cow
- Entrez Gene: 5604 Human
- Entrez Gene: 26395 Mouse
- Entrez Gene: 170851 Rat
- Omim: 176872 Human
- SwissProt: Q02750 Human
- SwissProt: P31938 Mouse
- SwissProt: Q01986 Rat
see all -
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
see all
Images
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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 - ab32091 observed at 43 kDa. Red - loading control, ab18058, observed at 130 kDa.
ab32091 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. Ab32091 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 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.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling MEK1 with purified ab32091 at dilution of 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
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Anti-MEK1 antibody [E342] (ab32091) at 1/5000 dilution + A431 (Human epidermoid carcinoma epithelial cell)whole cell lysate at 20 µg
Secondary
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?
Exposure time: 15 secondsBlocking/Diluting buffer and concentration: 5% NFDM /TBST
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Ab32091, at a 1/100 dilution, staining MEK1 in paraffin embedded human cervical carcinoma tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Overlay histogram showing HeLa cells stained with ab32091 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32091, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (43)
ab32091 has been referenced in 43 publications.
- Peng M et al. Programmed death-ligand 1 signaling and expression are reversible by lycopene via PI3K/AKT and Raf/MEK/ERK pathways in tongue squamous cell carcinoma. Genes Nutr 17:3 (2022). PubMed: 35164673
- Dong Y et al. Solasonine Induces Apoptosis and Inhibits Proliferation of Bladder Cancer Cells by Suppressing NRP1 Expression. J Oncol 2022:7261486 (2022). PubMed: 35281516
- Dessauges C et al. Optogenetic actuator - ERK biosensor circuits identify MAPK network nodes that shape ERK dynamics. Mol Syst Biol 18:e10670 (2022). PubMed: 35694820
- Bai D et al. BPTF activates the MAPK pathway through coexpression with Raf1 to promote proliferation of T-cell lymphoma. Oncol Lett 24:223 (2022). PubMed: 35720479
- Xie C et al. Identification of a BRAF/PA28γ/MEK1 signaling axis and its role in epithelial-mesenchymal transition in oral submucous fibrosis. Cell Death Dis 13:701 (2022). PubMed: 35961969