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

Overview

  • Product nameJNK 1/2/3 Total PhosphoTracer ELISA Kit
  • Detection methodFluorescent
  • Tests
    1 x 96 well plate
  • Sample type
    Cell culture extracts
  • Assay typeSemi-quantitative
  • Sensitivity
    5 ng/ml
  • Range
    5 ng/ml - 5000 ng/ml
  • Assay time
    2h 0m
  • Assay durationOne step assay
  • 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
  • PlatformMicroplate

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 1
    • c-Jun N-terminal kinase 2
    • c-Jun N-terminal kinase 3
    • JNK
    • JNK-46
    • JNK-55
    • JNK1
    • JNK1A2
    • JNK2
    • JNK21B1/2
    • JNK2A
    • JNK2ALPHA
    • JNK2B
    • JNK2BETA
    • JNK3
    • JNK3A
    • MAP kinase 10
    • MAP kinase 8
    • MAP kinase 9
    • MAP kinase p49 3F12
    • MAPK 10
    • MAPK 8
    • MAPK 9
    • Mapk9
    • Mitogen-activated protein kinase 10
    • Mitogen-activated protein kinase 8
    • Mitogen-activated protein kinase 9
    • MK09_HUMAN
    • p493F12
    • p54a
    • p54aSAPK
    • p54bSAPK
    • PRKM10
    • PRKM8
    • PRKM9
    • SAPK
    • SAPK1
    • SAPK1a
    • SAPK1b
    • SAPK1c
    • Stress-activated protein kinase 1a
    • Stress-activated protein kinase 1b
    • Stress-activated protein kinase 1c
    • Stress-activated protein kinase JNK1
    • Stress-activated protein kinase JNK2
    • Stress-activated protein kinase JNK3
    see all
  • Database links

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 Use at an assay dependent concentration. Tested in HEK293, HeLa, A431, C2C12, Raw264.7, A549, PC3, NIH3T3, U937

JNK 1/2/3 Total PhosphoTracer ELISA Kit images

  • 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.
  • 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.
  • 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.
  • PhosphoTracer JNK 1/2/3 Total ELISA Kit (ab119635) used in Sandwich ELISA.

Protocols

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

ab119635 has not yet been referenced specifically in any publications.

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

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

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"