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p38 MAPK alpha (Thr180/Tyr182) In-Cell ELISA Kit (ab126425)

Q&A (1)

Overview

  • Product namep38 MAPK alpha (Thr180/Tyr182) In-Cell ELISA Kit
  • Tests
    1 x 96 well plate
  • Sample type
    Adherent cells
  • Assay typeCell-based
  • Assay time
    5h 10m
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Product overview

    ab126425 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 p38 MAPK (Thr180/Tyr182) 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 p38 MAPK 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 p38 MAPK (Thr180/Tyr182) 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-p38 MAPK (Thr180/Tyr182) or Anti-p38 MAPK (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 applicationsIn-Cell ELISA more details

Properties

  • Storage instructionsStore at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    Anti-Mouse IgG Concentrate (Item I) 1 x 10µl
    Blocking Solution (5x) (Item F) 1 x 20ml
    Fixing Solution (Item D) 1 x 30ml
    Microplate (Item A) 1 unit
    Mouse Anti-p38 MAPK Concentrate (Item H) 1 x 7µl
    Mouse Anti-Phospho-p38 MAPK (Thr180/Tyr182) Concentrate (Item G) 1 x 7µl
    Quenching Buffer Concentrate (Item E) 1 x 2ml
    Stop Solution (Item K) 1 x 14ml
    TMB One-Step Substrate Reagent (Item J) 1 x 12ml
    Wash Buffer Concentrate A (20x) (Item B) 1 x 30ml
    Wash Buffer Concentrate B (20x) (Item C) 1 x 30ml
  • Research Areas
  • FunctionResponds to activation by environmental stress, pro-inflammatory cytokines and lipopolysaccharide (LPS) by phosphorylating a number of transcription factors, such as ELK1 and ATF2 and several downstream kinases, such as MAPKAPK2 and MAPKAPK5. Plays a critical role in the production of some cytokines, for example IL-6. May play a role in stabilization of EPO mRNA during hypoxic stress. Isoform Mxi2 activation is stimulated by mitogens and oxidative stress and only poorly phosphorylates ELK1 and ATF2. Isoform Exip may play a role in the early onset of apoptosis.
  • Tissue specificityBrain, heart, placenta, pancreas and skeletal muscle. Expressed to a lesser extent in lung, liver and kidney.
  • 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-180 and Tyr-182, which activates the enzyme.
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localizationCytoplasm. Nucleus.

    • Target information above from: UniProt accession Q16539 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
    • Alternative names
        CSAID-binding proteinCsaids binding proteinCSBP
        CSBP1CSBP2CSPB1Cytokine suppressive anti-inflammatory drug-binding proteinEXIPMAP kinase 14MAP kinase MXI2MAP kinase p38 alphaMAPK 14MAPK14MAX-interacting protein 2Mitogen-activated protein kinase 14Mitogen-activated protein kinase p38 alphaMK14_HUMANMxi2p38p38 MAP kinasep38 mitogen activated protein kinasep38alphap38alpha ExipPRKM14PRKM15RKSAPK2AStress activated protein kinase 2A
      see all
  • Database links

      • Applications

      Our Abpromise guarantee covers the use of ab126425 in the following tested applications.

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

      Application Notes
      In-Cell ELISA In-Cell ELISA

      p38 MAPK alpha (Thr180/Tyr182) In-Cell ELISA Kit images

      • In-Cell ELISA - p38 MAPK (Thr180/Tyr182) In-Cell ELISA Kit (ab126425)
        In-Cell ELISA - p38 MAPK (Thr180/Tyr182) In-Cell ELISA Kit (ab126425)
        HeLa cells were stimulated by different concentrations of anisomycin for 15 minutes at 37°C.
      • In-Cell ELISA - p38 MAPK (Thr180/Tyr182) In-Cell ELISA Kit (ab126425)
        In-Cell ELISA - p38 MAPK (Thr180/Tyr182) In-Cell ELISA Kit (ab126425)
        HeLa cells were stimulated by different concentrations of anisomycin for 1 hour at 37°C.
      • Western blot - p38 MAPK (Thr180/Tyr182) In-Cell ELISA Kit (ab126425)
        Western blot - p38 MAPK (Thr180/Tyr182) In-Cell ELISA Kit (ab126425)
        Western blot analysis of extracts from 1 µg/ml Anisomycin treated HeLa cells. Phospho-p38 MAPK (Thr180/Tyr182) and Anti-p38 MAPK antibodies from ab126425 were used in both detection assays.

      Protocols

      References for p38 MAPK alpha (Thr180/Tyr182) In-Cell ELISA Kit (ab126425)

      ab126425 has not yet been referenced specifically in any publications.

      Publishing research using ab126425 ? Please let us know so that we can cite the reference in this datasheet

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      I have a question regarding the kit p38 MAPK alpha (Thr180/Tyr182) In-Cell ELISA Kit (ab126425). Does this kit specifically
      identify the alpha isoform (either phosphorylated or total)? because from the component list, the antibodies seem to iden...

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      I'm sorry for the confusion. p38 is also know as 'Mitogen-activated protein kinase p38 alpha'. The term 'alpha' in the title of the product does not refer to an isoform of p38. p38 does have a few different isoforms but as far as I'm aware they ...

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      Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"