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Overview

  • Product nameAkt (pS473) ELISA KitSee all AKT 1/2/3 (phospho-Ser473) kits ...
  • Tests
    1 x 96 well plate
  • Sample type
    Cell Lysate
  • Assay typeSandwich
  • Assay time
    5h 00m
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Product overview

    ab126432 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human, mouse and rat cell lysates. By determining phosphorylated Akt protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

    This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human, mouse and rat phospho-Akt (Ser473). An anti-pan Akt antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and Akt present in a sample is bound to the wells by the immobilized antibody. The wells are washed and anti-Akt (Ser473) antibody is used to detect phosphorylated Akt (Ser473). After washing away unbound antibody, HRP-conjugated anti-rabbit IgG is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Akt (Ser473) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

  • Tested applicationsSandwich ELISA more details

Properties

  • Storage instructionsStore at -20°C. Please refer to protocols.
  • Components Identifier 1 x 96 tests
    20X Wash Buffer Concentrate Item B 1 x 25ml
    2X Cell Lysate Buffer Item J 1 x 5ml
    Akt Microplate (12 strips x 8 wells) coated with anti-pan Akt antibody 1 unit
    Assay Diluent: 5x concentrated buffer 1 x 15ml
    Detection Antibody Akt (Ser473): rabbit anti-phospho-Akt (Ser473) 2 vials
    HRP-conjugated anti-rabbit IgG: 1000x HRP-conjugated anti-rabbit IgG concentrate 1 x 25µl
    Positive Control: lyophilized powder from A431 cell lysate 1 vial
    Stop Solution Item I 1 x 8ml
    TMB One-Step Substrate Reagent Item H 1 x 12ml
  • Research Areas
  • FunctionGeneral protein kinase capable of phosphorylating several known proteins.
  • Tissue specificityExpressed in all human cell types so far analyzed.
  • Sequence similaritiesBelongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. RAC subfamily.
    Contains 1 AGC-kinase C-terminal domain.
    Contains 1 PH domain.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications    Phosphorylation on Thr-309 and Ser-474 is required for full activity.
    Ubiquitinated; undergoes both 'Lys-48'- and 'Lys-63'-linked polyubiquitination. TRAF6-induced 'Lys-63'-linked AKT2 ubiquitination. When fully phosphorylated and translocated into the nucleus, undergoes 'Lys-48'-polyubiquitination catalyzed by TTC3, leading to its degradation by the proteasome.

    • Target information above from: UniProt accession P31751 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
    • Alternative names
        Akt2AKT2_HUMANPKB beta
        Protein kinase Akt-2Protein kinase B betaRAC-beta serine/threonine-protein kinaseRAC-PK-beta
      see all
  • Database links
    see all

    • Applications

    Our Abpromise guarantee covers the use of ab126432 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

    Akt (pS473) ELISA Kit images

    • Sandwich ELISA - Akt (pSer473) ELISA Kit (ab126432)
      Sandwich ELISA - Akt (pSer473) ELISA Kit (ab126432)
      A431 cells were treated with recombinant human EGF at 37°C for 20 min. Solubilize cells at 4 x 107 cells/ml in Cell Lysate Buffer. Serial dilutions of lysates were analyzed in this ELISA.
    • Sandwich ELISA - Akt (pSer473) ELISA Kit (ab126432)
      Sandwich ELISA - Akt (pSer473) ELISA Kit (ab126432)
      NIH3T3 cells were treated or untreated with recombinant human PDGF for 10 min. Cell lysates were analyzed using this phosphoELISA.
    • Western blot - Akt (pSer473) ELISA Kit (ab126432)
      Western blot - Akt (pSer473) ELISA Kit (ab126432)
      NIH3T3 cells were treated or untreated with recombinant human PDGF for 10 min. Cell lysates were analyzed by Western Blot.
    • Sandwich ELISA - Akt (pSer473) ELISA Kit (ab126432)
      Sandwich ELISA - Akt (pSer473) ELISA Kit (ab126432)
      The NIH3T3 cells were treated with recombinant human PDGF for 10 minutes to induce phosphorylation of Akt. Serial dilutions of lysates were analyzed in this ELISA.

    Protocols

    References for Akt (pS473) ELISA Kit (ab126432)

    ab126432 has not yet been referenced specifically in any publications.

    Publishing research using ab126432 ? Please let us know so that we can cite the reference in this datasheet

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