Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y78] to MEK2
- Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
- Knockout validated
- Reacts with: Mouse, Human
Product nameAnti-MEK2 antibody [Y78]
See all MEK2 primary antibodies
DescriptionRabbit monoclonal [Y78] to MEK2
SpecificityThis antibody does not cross react with other MAP kinase kinase family members
Tested applicationsSuitable for: ICC/IF, WB, IHC-P, Flow Cytmore details
Species reactivityReacts with: Mouse, Human
Synthetic peptide. within Human MEK2 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P36507
- WB: Jurkat whole cell lysate (ab7899). ICC/IF: HeLa and wildtype HAP1 cells. IHC-P: Human prostate carcinoma tissue.
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
- Anti-MEK2 antibody [Y78] (Alexa Fluor® 647) (ab200519)
- Anti-MEK2 antibody [Y78] (Alexa Fluor® 488) (ab200606)
- Anti-MEK2 antibody [Y78] (HRP) (ab200607)
- Anti-MEK2 antibody [Y78] (PE) (ab213661)
- Anti-MEK2 antibody [Y78] (Alexa Fluor® 555) (ab214633)
- Anti-MEK2 antibody [Y78] (Alexa Fluor® 594) (ab214845)
- Anti-MEK2 antibody [Y78] - BSA and Azide free (ab233731)
Our Abpromise guarantee covers the use of ab32517 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 1 µg/ml.|
|WB||1/10000. Detects a band of approximately 45 kDa (predicted molecular weight: 44 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.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionCatalyzes the concomitant phosphorylation of a threonine and a tyrosine residue in a Thr-Glu-Tyr sequence located in MAP kinases. Activates the ERK1 and ERK2 MAP kinases.
Involvement in diseaseDefects in MAP2K2 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 similaritiesBelongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase subfamily.
Contains 1 protein kinase domain.
modificationsMAPKK is itself dependent on Ser/Thr phosphorylation for activity catalyzed by MAP kinase kinase kinases (RAF or MEKK1).
Acetylation of Ser-222 and Ser-226 by Yersinia yopJ prevents phosphorylation and activation, thus blocking the MAPK signaling pathway.
- Information by UniProt
- Cardiofaciocutaneous syndrome antibody
- CFC syndrome antibody
- CFC4 antibody
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MEK2 knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: K562 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab32517 observed at 44 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32517 was shown to specifically react with MEK2 when MEK2 knockout samples were used. Wild-type and MEK2 knockout samples were subjected to SDS-PAGE. ab32517 and ab8245 (loading control to GAPDH) were diluted 1/10 000 and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
ab32517 staining MEK2 in wild-type HAP1 cells (top panel) and MEK2 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32517 at 1μg/ml and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
All lanes : Anti-MEK2 antibody [Y78] (ab32517) at 1/10000 dilution
Lane 1 : Jurkat Whole Cell Lysate
Lane 2 : Mouse Brain Tissue Lysate
Lane 3 : Mouse Lung Tissue Lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 44 kDa
Observed band size: 44 kDa
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab32517 overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) at a 1:10000 dilution for 1hr at room temperature and then imaged.
Anti-MEK2 antibody [Y78] (ab32517) at 1/10000 dilution + Jurkat cell lysate
Predicted band size: 44 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Immunofluorescent staining of HeLa cells using ab32517 at a dilution of 1/250.
Immunohistochemical analysis of paraffin embedded human prostate carcinoma using ab32517 at a dilution of 1/250.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Overlay histogram showing HeLa cells stained with ab32517 (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 (ab32517, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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.
ab32517 has been referenced in 6 publications.
- An XZ et al. Netrin-1 suppresses the MEK/ERK pathway and ITGB4 in pancreatic cancer. Oncotarget 7:24719-33 (2016). PubMed: 27034160
- Li L et al. MEK1 promotes YAP and their interaction is critical for tumorigenesis in liver cancer. FEBS Lett 587:3921-7 (2013). WB . PubMed: 24211253
- Maïga O et al. Identification of mitogen-activated protein/extracellular signal-responsive kinase kinase 2 as a novel partner of the scaffolding protein human homolog of disc-large. FEBS J 278:2655-65 (2011). PubMed: 21615688
- Yamashita S et al. The Leydig cell MEK/ERK pathway is critical for maintaining a functional population of adult Leydig cells and for fertility. Mol Endocrinol 25:1211-22 (2011). WB ; Mouse . PubMed: 21527500
- Zhu H et al. EGFR signals downregulate tumor suppressors miR-143 and miR-145 in Western diet-promoted murine colon cancer: role of G1 regulators. Mol Cancer Res 9:960-75 (2011). WB . PubMed: 21653642
- Nadeau V et al. Map2k1 and Map2k2 genes contribute to the normal development of syncytiotrophoblasts during placentation. Development 136:1363-74 (2009). PubMed: 19304888