Overview

  • Product name

    Anti-JNK2 antibody [EP1595Y] - BSA and Azide free
    See all JNK2 primary antibodies
  • Description

    Rabbit monoclonal [EP1595Y] to JNK2 - BSA and Azide free
  • Host species

    Rabbit
  • 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

    Ab227986 is the carrier-free version of ab76125. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab227986 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab227986 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 Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), 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

  • This WB data was generated using the same anti-JNK2 antibody clone, EP1595Y, in a different buffer formulation (cat# ab76125).

    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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76125).

  • 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 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76125).

  • This IHC data was generated using the same anti-JNK2 antibody clone, EP1595Y, in a different buffer formulation (cat# ab76125).

    ab76125 at 1/100 dilution staining JNK2 in human breast carcinoma by Immunohistochemistry, Paraffin-embedded tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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) »
  • 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) »
See all 10 Publications for this product

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