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Akt (pS473) + total Akt ELISA Kit (ab126433)

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Overview

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

    ab126433 is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in 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 Blot analysis.

    This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of phospho-Akt (Ser473) and total Akt in human, mouse and rat cell lysates (help normalize the results of phospho-Akt from different cell lysate being compared). A 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-phospho-Akt (Ser473) or anti-pan-Akt is used to detect phosphorylated or total Akt. 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) or total Akt 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
  • PlatformMicroplate

Properties

  • Storage instructionsStore at -20°C. Please refer to protocols.
  • Components 1 x 96 tests
    20X Wash Buffer Concentrate 1 x 25ml
    2X Cell Lysate Buffer 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) 1 vial
    Detection Antibody Akt: rabbit anti-pan-Akt 1 vial
    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 1 x 8ml
    TMB One-Step Substrate Reagent 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
        AKTAKT1Akt2
        AKT2_HUMANAKT3HIHGHHPKBPKB alphaPKB betaPKB gammaPKB-ALPHAPKB-GAMMAPKBBPKBBETAPKBGPRKBAPRKBBPRKBGProtein kinase Akt-2Protein kinase Akt-3Protein kinase BProtein kinase B alphaProtein kinase B betaProtein kinase B gammaProto oncogene c AktRACRAC-ALPHARAC-alpha serine/threonine-protein kinaseRAC-BETARAC-beta serine/threonine-protein kinaseRAC-gammaRAC-gamma serine/threonine-protein kinaseRAC-PK-alphaRAC-PK-betaRAC-PK-gammaSTK-2V-akt murine thymoma viral oncogene homolog 1V-akt murine thymoma viral oncogene homolog 2V-akt murine thymoma viral oncogene homolog 3 (protein kinase B, gamma)
      see all
  • Database links
    see all

    • Applications

    Our Abpromise guarantee covers the use of ab126433 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) + total Akt ELISA Kit images

    • Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)
      Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)
      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 (pS473) + total Akt ELISA Kit (ab126433)
      Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)
      NIH3T3 cells were treated or untreated with recombinant human PDGF for 10 min. Cell lysates were analyzed using this phosphoELISA.
    • Western blot - Akt (pS473) + total Akt ELISA Kit (ab126433)
      Western blot - Akt (pS473) + total Akt ELISA Kit (ab126433)
      NIH3T3 cells were treated or untreated with recombinant human PDGF for 10 min. Cell lysates were analyzed by Western Blot.
    • Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)
      Sandwich ELISA - Akt (pS473) + total Akt ELISA Kit (ab126433)
      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) + total Akt ELISA Kit (ab126433)

    ab126433 has not yet been referenced specifically in any publications.

    Publishing research using ab126433 ? Please let us know so that we can cite the reference in this datasheet

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    Please clarify if ab126433 measures both Akt (pS473) and total Akt simultaneously in the same well?


    I can confirm that this kit uses a plate coated with pan Akt antibody. After samples have been added, either anti-phospho-Akt (Ser473) or anti-pan-Akt antibody is used to detect phosphorylated or total Akt. This means that either Akt (pS473) or total...

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    we would be interested in the pAkt ELISA Kit ab126433, that is out of
    stock. Is it still possible to order?
    I have got another question regarding quantitative analysis of
    extracellular matrix components of tissue fragments. Which method

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    Thank you for contacting us.


    I am happy to confirm that ab126433 is in stock in our US office and if you order it today before 4pm you should receive it on the 3rd of July.


    We are happy to help customers find the most suita...

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    Product code: 126433
    Inquiry: The customer wants to buy the Akt (pS473) + total Akt ELISA Kit (ab126433) .But the kits is out of stock .He wants to know when he can get the kits. Would you please help him provide the information? Thank you very much.

    Thank you for contacting us.

    We have just received a shipment into stock (this informationwill be updated on the website shortly) and will be receiving more in the future. If your customer were to place an order we would be able to send them out...

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