Overview

  • Product name
    Anti-JNK2 antibody [EP1595Y]
    See all JNK2 primary antibodies
  • Description
    Rabbit monoclonal [EP1595Y] to JNK2
  • Tested applications
    Suitable for: WB, IP, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues near the C terminus of human JNK2.

  • Positive control
    • HeLa cell lysate; human breast carcinoma tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    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

Properties

Applications

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.

Application Abreviews Notes
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.
Flow Cyt 1/40. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Responds 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 similarities
    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • Domain
    The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • Post-translational
    modifications
    Dually phosphorylated on Thr-183 and Tyr-185, which activates the enzyme. Autophosphorylated in vitro.
  • Information by UniProt
  • Database links
  • Alternative names
    • c Jun kinase 2 antibody
    • C Jun N terminal kinase 2 antibody
    • c-Jun N-terminal kinase 2 antibody
    • JNK 55 antibody
    • JNK-55 antibody
    • JNK2 alpha antibody
    • JNK2 antibody
    • JNK2 beta antibody
    • JNK2A antibody
    • JNK2alpha antibody
    • JNK2B antibody
    • JNK2BETA antibody
    • Jun kinase antibody
    • MAP kinase 9 antibody
    • MAPK 9 antibody
    • Mapk9 antibody
    • Mitogen activated protein kinase 9 antibody
    • Mitogen-activated protein kinase 9 antibody
    • MK09_HUMAN antibody
    • P54a antibody
    • p54aSAPK antibody
    • PRKM9 antibody
    • Protein kinase, mitogen-activated, 9 antibody
    • SAPK alpha antibody
    • SAPK antibody
    • SAPK1a antibody
    • Stress activated protein kinase 1a antibody
    • Stress-activated protein kinase JNK2 antibody
    see all

Images



  • Predicted band size : 48 kDa

    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

    Secondary
    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.

References

This product has been referenced in:
  • Kitanaka T  et al. JNK activation is essential for activation of MEK/ERK signaling in IL-1ß-induced COX-2 expression in synovial fibroblasts. Sci Rep 7:39914 (2017). RT-PCR ; Cat . Read more (PubMed: 28054591) »
  • Wang B  et al. IL-34 Upregulated Th17 Production through Increased IL-6 Expression by Rheumatoid Fibroblast-Like Synoviocytes. Mediators Inflamm 2017:1567120 (2017). WB ; Human . Read more (PubMed: 28659662) »

See all 10 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Cat Cell lysate - whole cell (synovial fibroblasts)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
10 µg
Treatment
siRNA for scramble control, JNK1 and JNK2
Specification
synovial fibroblasts
Blocking step
(agent) for 50 minute(s) · Concentration: 100% · Temperature: 25°C
Username

令 中野

Verified customer

Submitted Jun 29 2017

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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