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PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635)

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Overview

  • Product namePhosphoTracer JNK 1/2/3 Total ELISA Kit
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSandwich
  • Sensitivity
    5 ng/ml
  • Range
    5 ng/ml - 5000 ng/ml
  • Assay time
    1h 15m
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Product overview

    PhosphoTracer assays use a traditional immuno-sandwich format, but with a major difference both the analyte and the assay reagents are added to the PhosphoTracer assay microplate at the same time. After a short incubation period, unbound assay reagents and analytes are washed away, and immuno-complexes containing both antibodies are detected. The process can take as little as 60 minutes to complete.

    PhosphoTracer kits also allow a higher degree of assay flexibility. In contrast to other ELISA formats, no antibodies are present on the assay microplate itself, so assays for several different targets can be performed in different wells on the same microplate. Simply mix the lysate with your target reagents of choice, using the microplate configuration of your choice.

    A whole new way of performing cellular assays, PhosphoTracer takes the hard work out of running a standard ELISA, while still giving the high quality results expected from a sandwich immunoassay. Fully self-contained kits are supplied in convenient 96-well packs. Simple to use and highly sensitive PhosphoTracer kits are designed to get results, fast.

     

    Abcam’s PhosphoTracer JNK 1/2/3 assay kits detect endogenous levels of JNK 1/2/3 (GenBank Accessions NP_620637 [JNK1], NP_620707 [JNK2] and NP_620446 [JNK3]) in cellular lysates. Total JNK 1/2/3 assay kits detect JNK irrespective of phosphorylation status.



    The substrate used with the HRP conjugated detection antibody is a combination of 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP) (wavelength exc/em = 530-540nm / 590-600nm), a highly sensitive and stable substrate for HRP) and ADHP Dilution Buffer (a stabilized H2O2 solution). Learn more about the fluorogenic substrate, ADHP.

  • Tested applicationsSandwich ELISA more details

Properties

  • 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.
  • Post-translational
    modifications    Dually phosphorylated on Thr-183 and Tyr-185, which activates the enzyme. Autophosphorylated in vitro.

    • Target information above from: UniProt accession P45984 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
    • Alternative names
        c-Jun N-terminal kinase 1c-Jun N-terminal kinase 2c-Jun N-terminal kinase 3
        JNKJNK-46JNK-55JNK1JNK1A2JNK2JNK21B1/2JNK2AJNK2ALPHAJNK2BJNK2BETAJNK3JNK3AMAP kinase 10MAP kinase 8MAP kinase 9MAP kinase p49 3F12MAPK 10MAPK 8MAPK 9Mapk9Mitogen-activated protein kinase 10Mitogen-activated protein kinase 8Mitogen-activated protein kinase 9MK09_HUMANp493F12p54ap54aSAPKp54bSAPKPRKM10PRKM8PRKM9SAPKSAPK1SAPK1aSAPK1bSAPK1cStress-activated protein kinase 1aStress-activated protein kinase 1bStress-activated protein kinase 1cStress-activated protein kinase JNK1Stress-activated protein kinase JNK2Stress-activated protein kinase JNK3
      see all
  • Database links
    see all

    • Applications

    Our Abpromise guarantee covers the use of ab119635 in the following tested applications.

    The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

    Application Notes
    Sandwich ELISA sELISA. Tested in HEK293, HeLa, A431, C2C12, Raw264.7, A549, PC3, NIH3T3, U937

    PhosphoTracer JNK 1/2/3 Total ELISA Kit images

    • Western blot - PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635)
      Western blot - PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635)
      Using Western Blot, a significant stimulation of JNK 1/2/3 phosphorylation at Thr183/Tyr185 is detected in HEK293 cells treated with anisomycin for 30 minutes (+) compared with untreated cells (-), while no change in total JNK levels is observed.
    • Sandwich ELISA - PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635)
      Sandwich ELISA - PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635)
      Using the JNK 1/2/3 assay kits, a significant stimulation of JNK 1/2/3 phosphorylation at Thr183/Tyr185 is detected in HEK293 cells treated with anisomycin for 30 minutes (+) compared with untreated cells (-), while no change in total JNK levels is observed.
    • Sandwich ELISA - PhosphoTracer JNK 1/2/3 Total ELISA kit (ab119635)
      Sandwich ELISA - PhosphoTracer JNK 1/2/3 Total ELISA kit (ab119635)
      A dilution series of recombinant active JNK1alpha was prepared by diluting to various concentrations in Lysis Mix, containing 0.1% BSA. These dilution series were analyzed using the PhosphoTracer phospho-JNK assay. Limit of detection of active rhJNK was around 10 ng/mL.
    • Sandwich ELISA - PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635)
      Sandwich ELISA - PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635)
      PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635) used in Sandwich ELISA.

    Protocols

    References for PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635)

    ab119635 has not yet been referenced specifically in any publications.

    Publishing research using ab119635 ? Please let us know so that we can cite the reference in this datasheet

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    Submit an Abreview for ab119635 Submit a Question for ab119635

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    10
    Votes: Highest First

    Dear Madam / Sir,

    Regarding Phosphotracer ELISA assays for JNK and IKBa, is there any experience using adherent monocytes?

    Thank you in advance for your help.

    Thank you for contacting us.

    These kits have not been tested in adherent monocytes but should be suitable for use in any cell extract.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any mo...

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    Kits phosphotracer
    cibles ERK 1/2, p38 MAPKalpha, JNK 1/2/3.

    Merci de nous avoir contactés.

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    ERK 1/2 (pThr202/Tyr204) : ab119625, http://www.abcam.com/ab119625
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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"