Overview

  • Product name

    p38 MAPK alpha ELISA Kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    HeLa extract 6 9.2%
    Inter-assay
    Sample n Mean SD CV%
    HeLa extract 3 6.4%
  • Sample type

    Cell Lysate, Tissue Homogenate
  • Assay type

    Semi-quantitative
  • Sensitivity

    300 pg/ml
  • Range

    0.781 ng/ml - 100 ng/ml
  • Assay time

    1h 1m
  • Assay duration

    One step assay
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Product overview

    p38 MAPK alpha ELISA Kit (ab176650) has been re-developed with new capture and detector antibodies. This new kit has the same name but a different product number (ab221012). We have identified new recombinant monoclonal antibodies to use in the SimpleStep ELISA platform that provide a higher sensitivity when quantifying p38 MAPKα protein in Human and mouse cells.


    Abcam’s p38 MAPKα in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of p38 MAPKα protein in Human and mouse cells.


    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    10X Wash Buffer PT 1 x 20ml
    50X Cell Extraction Enhancer Solution 1 x 1ml
    5X Cell Extraction Buffer PTR 1 x 12ml
    Lyophilized p38 MAPK alpha Control Lysate 1 vial
    p38 MAPK alpha (Total) Capture Antibody 1 x 3ml
    p38 MAPK alpha (Total) Detector Antibody 1 x 3ml
    Plate Seal 1 unit
    SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit
    Stop Solution 1 x 12ml
    TMB Substrate 1 x 12ml
  • 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
    • 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 ab221012 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.

     

  • Raw data values are fitted to a four-parameter, variable slope curve (+/- SD).

  • Raw data values are fitted to a four-parameter, variable slope curve (+/- SD).

  • Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

  • HeLa cells were cultured in 96-well tissue culture plates and treated (15 min) with a dose-range of anisomycin before cell lysis. Data from quadruplicate measurements of p38 MAPKα (pT180/Y182) are plotted and compared against Total p38 MAPKα protein levels.

Protocols

References

ab221012 has not yet been referenced specifically in any publications.

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