Overview

  • Product name
    JNK (Thr183/Tyr185) In-Cell ELISA Kit
  • Detection method
    Colorimetric
  • Sample type
    Adherent cells
  • Assay type
    Cell-based (qualitative)
  • Assay time
    5h 10m
  • Assay duration
    Multiple steps standard assay
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Product overview

    ab126424 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in cells. It can be used for measuring the relative amount of JNK (Thr183/Tyr185) phosphorylation and screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators in cultured Human, Mouse and Rat cell lines. By determining JNK protein phosphorylation in your experimental model system, you can verify pathway activation in your cell lines without spending excess time and effort in preparing cell lysate and performing an analysis of Western Blot.


    In the JNK (Thr183/Tyr185) In-Cell ELISA Kit, cells are seeded into a 96 well tissue culture plate. The cells are fixed after various treatments, inhibitors or activators. After blocking, anti-Phospho-JNK(Thr183/Tyr185) or anti-JNK (primary antibody) is pipetted into the wells and incubated. The wells are washed, and HRP-conjugated anti-Mouse IgG (secondary antibody) is added to the wells. The wells are washed again, a


    TMB substrate solution is added to the wells and color develops in proportion to the amount of protein. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Tested applications
    Suitable for: In-Cell ELISAmore details
  • Platform
    Microplate

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    HRP-conjugated Anti-Mouse IgG Concentrate 1 x 10µl
    Blocking Buffer Concentrate (5X) 1 x 20ml
    Fixing Solution 1 x 30ml
    Uncoated 96-well Microplate 1 unit
    Mouse anti-JNK Concentrate (Item H) 1 x 7µl
    Mouse anti-Phospho-JNK (Thr183/Tyr185) Concentrate (Item G) 1 x 7µl
    Quenching Buffer Concentrate (30x) 1 x 2ml
    Stop Solution 1 x 14ml
    TMB One-Step Substrate Reagent 1 x 12ml
    Wash Buffer A Concentrate (20X) 1 x 30ml
    Wash Buffer B Concentrate (20X) 1 x 30ml
  • Research areas
  • Function
    Serine/threonine-protein kinase involved in various processes such as cell proliferation, differentiation, migration, transformation and programmed cell death. Extracellular stimuli such as proinflammatory cytokines or physical stress stimulate the stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) signaling pathway. In this cascade, two dual specificity kinases MAP2K4/MKK4 and MAP2K7/MKK7 phosphorylate and activate MAPK8/JNK1. In turn, MAPK8/JNK1 phosphorylates a number of transcription factors, primarily components of AP-1 such as JUN, JDP2 and ATF2 and thus regulates AP-1 transcriptional activity. Phosphorylates the replication licensing factor CDT1, inhibiting the interaction between CDT1 and the histone H4 acetylase HBO1 to replication origins. Loss of this interaction abrogates the acetylation required for replication initiation. Promotes stressed cell apoptosis by phosphorylating key regulatory factors including p53/TP53 and Yes-associates protein YAP1. In T-cells, MAPK8 and MAPK9 are required for polarized differentiation of T-helper cells into Th1 cells. Contributes to the survival of erythroid cells by phosphorylating the antagonist of cell death BAD upon EPO stimulation. Mediates starvation-induced BCL2 phosphorylation, BCL2 dissociation from BECN1, and thus activation of autophagy. Phosphorylates STMN2 and hence regulates microtubule dynamics, controlling neurite elongation in cortical neurons. In the developing brain, through its cytoplasmic activity on STMN2, negatively regulates the rate of exit from multipolar stage and of radial migration from the ventricular zone. Phosphorylates several other substrates including heat shock factor protein 4 (HSF4), the deacetylase SIRT1, ELK1, or the E3 ligase ITCH.
    JNK1 isoforms display different binding patterns: beta-1 preferentially binds to c-Jun, whereas alpha-1, alpha-2, and beta-2 have a similar low level of binding to both c-Jun or ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms.
  • 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 by MAP2K7 and MAP2K4, which activates the enzyme. Phosphorylated by TAOK2.
  • Cellular localization
    Cytoplasm. Nucleus.
  • Information by UniProt
  • Alternative names
    • C Jun kinase 2
    • c Jun N terminal kinase 1
    • c Jun N terminal kinase 2
    • c Jun N terminal kinase 3
    • c-Jun N-terminal kinase 1
    • JNK
    • JNK 46
    • JNK 55
    • JNK-46
    • JNK1
    • JNK1A2
    • JNK2
    • JNK21B1/2
    • JNK2A
    • JNK2ALPHA
    • JNK2B
    • JNK2BETA
    • JNK3
    • JNK3 alpha protein kinase
    • JNK3A
    • Jun kinase
    • JUN N terminal kinase
    • MAP kinase 10
    • MAP kinase 8
    • MAP kinase 9
    • MAP kinase p49 3F12
    • MAPK 10
    • MAPK 8
    • MAPK 9
    • MAPK10
    • mapk8
    • MAPK9
    • Mitogen activated protein kinase 10
    • Mitogen activated protein kinase 8
    • Mitogen activated protein kinase 8 isoform JNK1 alpha1
    • Mitogen activated protein kinase 8 isoform JNK1 beta2
    • Mitogen activated protein kinase 9
    • Mitogen-activated protein kinase 8
    • MK08_HUMAN
    • p493F12
    • p54a
    • p54aSAPK
    • p54bSAPK
    • PRKM10
    • PRKM8
    • PRKM9
    • SAPK
    • SAPK(beta)
    • SAPK1
    • SAPK1a
    • SAPK1b
    • SAPK1c
    • Stress activated protein kinase 1
    • Stress activated protein kinase 1a
    • Stress activated protein kinase 1b
    • Stress activated protein kinase 1c
    • Stress activated protein kinase beta
    • Stress activated protein kinase JNK1
    • Stress activated protein kinase JNK2
    • Stress activated protein kinase JNK3
    • Stress-activated protein kinase 1c
    • Stress-activated protein kinase JNK1
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab126424 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
In-Cell ELISA Use at an assay dependent concentration.

Images

  • HeLa cells were stimulated by different concentrations of anisomycin for 15 minutes at 37°C.

     

     

  • Western blot analysis of extracts from 1 µg/ml Anisomycin treated HeLa cells. Anti-Phospho-JNK (Thr183/Tyr185) and Anti-JNK antibodies were used in both detection assays.
  • HeLa cells were stimulated by different concentrations of anisomycin for 15 minutes at 37°C.
  • HeLa cells were stimulated by different concentrations of anisomycin for 1 hour at 37°C.

Protocols

References

This product has been referenced in:
  • Behera J  et al. Hydrogen sulfide epigenetically mitigates bone loss through OPG/RANKL regulation during hyperhomocysteinemia in mice. Bone 114:90-108 (2018). Read more (PubMed: 29908298) »
See 1 Publication for this product

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