Product nameAnti-JNK2 antibody [EP1595Y]
See all JNK2 primary antibodies
DescriptionRabbit monoclonal [EP1595Y] to JNK2
Tested applicationsSuitable for: WB, IP, IHC-P, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human JNK2 aa 350-450 (C terminal). The exact sequence is proprietary.
- HeLa whole cell lysate (ab150035); human breast carcinoma tissue.
A trial size is available to purchase for this antibody.
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.5% BSA
Concentration information loading...
PurityProtein A purified
Our Abpromise guarantee covers the use of ab76125 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 48 kDa.|
|IP||Use a concentration of 5 µg/ml.|
|IHC-P||1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionResponds to activation by environmental stress and pro-inflammatory cytokines by phosphorylating a number of transcription factors, primarily components of AP-1 such as c-Jun and ATF2 and thus regulates AP-1 transcriptional activity. In T-cells, JNK1 and JNK2 are required for polarized differentiation of T-helper cells into Th1 cells.
JNK2 isoforms display different binding patterns: alpha-1 and alpha-2 preferentially bind to c-Jun, whereas beta-1 and beta-2 bind to ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms. JUNB is not a substrate for JNK2 alpha-2, and JUND binds only weakly to it.
Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain.
DomainThe TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
modificationsDually phosphorylated on Thr-183 and Tyr-185, which activates the enzyme. Autophosphorylated in vitro.
- Information by UniProt
- c Jun kinase 2 antibody
- C Jun N terminal kinase 2 antibody
- c-Jun N-terminal kinase 2 antibody
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: JNK2 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: MCF7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab76125 observed at 54 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab76125 was shown to specifically react with JNK2 when JNK2 knockout samples were used. Wild-type and JNK2 knockout samples were subjected to SDS-PAGE. ab76125 and ab8245 (loading control to GAPDH) were diluted 1/2500 and 1/2000 respectively 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 1/10000 dilution for 1 h at room temperature before imaging.
JNK2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit monoclonal to JNK2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab76125.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 48kDa; JNK2
ab76125 at 1/100 dilution staining JNK2 in human breast carcinoma by Immunohistochemistry, Paraffin-embedded tissue.
Anti-JNK2 antibody [EP1595Y] (ab76125) at 1/50000 dilution + HeLa cell lysate at 10 µg
goat anti-rabbit-HRP at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 48 kDa
Observed band size: 54 kDa why is the actual band size different from the predicted?
Additional bands at: 46 kDa (possible isoform)
Overlay histogram showing HeLa cells stained with ab76125 (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 (ab76125, 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.
This product has been referenced in:
- Zhu Q et al. Cepharanthine exerts antitumor activity on choroidal melanoma by reactive oxygen species production and c-Jun N-terminal kinase activation. Oncol Lett 13:3760-3766 (2017). Read more (PubMed: 28529590) »
- Sheng X et al. Bacterial effector NleL promotes enterohemorrhagic E. coli-induced attaching and effacing lesions by ubiquitylating and inactivating JNK. PLoS Pathog 13:e1006534 (2017). Read more (PubMed: 28753655) »