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

Overview

  • Product name
    p38 MAPK alpha (Thr180/Tyr182) 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

    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 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
    Anti-Mouse IgG Concentrate (Item I) 1 x 10µl
    5X Blocking Buffer Concentrate (Item F) 1 x 20ml
    Fixing Solution (Item D) 1 x 30ml
    Uncoated 96-well 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 (30x) (Item E) 1 x 2ml
    Stop Solution 1 x 14ml
    TMB Substrate Reagent 1 x 12ml
    20X Wash Buffer A Concentrate (Item B) 1 x 30ml
    20X Wash Buffer B Concentrate (Item C) 1 x 30ml
  • Research areas
  • Function
    Responds 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 specificity
    Brain, heart, placenta, pancreas and skeletal muscle. Expressed to a lesser extent in lung, liver and kidney.
  • 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-180 and Tyr-182, which activates the enzyme.
    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization
    Cytoplasm. Nucleus.
  • Information by UniProt
  • Alternative names
    • CSAID-binding protein
    • Csaids binding protein
    • CSBP
    • CSBP1
    • CSBP2
    • CSPB1
    • Cytokine suppressive anti-inflammatory drug-binding protein
    • EXIP
    • MAP kinase 14
    • MAP kinase MXI2
    • MAP kinase p38 alpha
    • MAPK 14
    • MAPK14
    • MAX-interacting protein 2
    • Mitogen-activated protein kinase 14
    • Mitogen-activated protein kinase p38 alpha
    • MK14_HUMAN
    • Mxi2
    • p38
    • p38 MAP kinase
    • p38 mitogen activated protein kinase
    • p38alpha
    • p38alpha Exip
    • PRKM14
    • PRKM15
    • RK
    • SAPK2A
    • Stress 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 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.

     

  • HeLa cells were stimulated by different concentrations of anisomycin for 1 hour at 37°C.

     

  • 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.
  • 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:
  • Campo GM  et al. Beta-arrestin-2 negatively modulates inflammation response in mouse chondrocytes induced by 4-mer hyaluronan oligosaccharide. Mol Cell Biochem 399:201-8 (2015). Read more (PubMed: 25318610) »

See 1 Publication for this product

Customer reviews and Q&As

This kit provided a cell-based ELISA, easy to use.
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Abcam user community

Verified customer

Submitted Sep 21 2017



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 th...

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

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