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PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit (ab119634)

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Overview

  • Product namePhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSandwich
  • Sensitivity
    50 ng/ml
  • Range
    50 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. Phospho-JNK 1/2/3 assay kits detect JNK only when phosphorylated at Thr183/Tyr185. 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 3c-Jun N-terminal kinase 2
        JNKJNK 46JNK 55JNK-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 JNK3Stress-activated protein kinase JNK2
      see all
  • Database links
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    • Applications

    Our Abpromise guarantee covers the use of ab119634 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 HeLa, HEK293, U937, NIH3T3

    PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit images

    • Western blot - PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit (ab119634)
      Western blot - PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit (ab119634)
      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 (pT183/Y185) ELISA Kit (ab119634)
      Sandwich ELISA - PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit (ab119634)
      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 (pT183/Y185) ELISA kit (ab119634)
      Sandwich ELISA - PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA kit (ab119634)
      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 50 ng/mL.
    • Sandwich ELISA - PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit (ab119634)
      Sandwich ELISA - PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit (ab119634)
      PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit (ab119634) used in Sandwich ELISA.

    Protocols

    References for PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit (ab119634)

    ab119634 has not yet been referenced specifically in any publications.

    Publishing research using ab119634 ? Please let us know so that we can cite the reference in this datasheet

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

    Merci de nous avoir contactés.

    Voici la liste des kits

    1) pour la détection d'une seule formed'une protéine particulière :

    ERK 1/2 (pThr202/Tyr204) : ab119625, http://www.abcam.com/ab119625
    ER...

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    Does PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA kit allows absolute quantitation of JNK using standard curves?          Thank you in advance for your help.  

    Thank you for contacting us.


    This particular assay kit detect JNK only when phosphorylated at Thr183/Tyr185. Total JNK 1/2/3 assay kits detect JNK irrespective of phosphorylation status. 

    To analyze the data, averaged counts are ca...

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    I have a last question regarding the PhosphoTracer JNK1/2/3 ELISA kit (ab119634):

    Why the kit does not use formaldehyde solution? Fixed cells are stable for several weeks, and you can perform ELISA assay when desired.

    Thank you for contacting Abcam.

    The PhosphoTracer JNK 1/2/3 (pT183/Y185) ELISA Kit has been optimized to be used on fresh cell lyatses and we do not know if the cross-linking that occurs during fixation would affect the assay.

    If you d...

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    Hello, we are interested in testing a battery of phosphorylation events in our model and are interested in your Phosphotracer line of ELISA kits. I just want to confirm that we can run multiple target experiments on a single plate from each kit. i.e. if w...

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    Thank you for contacting Abcam regarding our Phosphotracer ELISA kits.

    I am happy to confirm that you can run multiple target experiments on each plate. The provided plates MUST be used but the reagents and assay principle are interchangeable. ...

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    Thank you very much for your answer.
    Unfortunately, it is very difficult to read the message because of a configuration problem in my email box.
    Could you send me your message at the following email addresse:

    Thank you in advance for ...

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    Thank you for contacting Abcam.

    Please find the answers to your questions below:

    1 - The expiration date will be on the product box when it arrives. However once the product is opened, it will remain viable for one month.

    2...

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    I'm interested in the product PhosphoTracer JNK1/2/3 ELISA kit (ab119634).

    I would like to know some aspects of this product:

    - The expiration time of every component of the kits

    - Intra- and interassay coefficients of var...

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    Thank you for contacting Abcam.

    Please find the answers to your questions below:
    1 - The expiration date will be on the product box when it arrives. However once the product is opened, it will remain viable for one month.
    2 - The Coeff...

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