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Signal Transduction Protein Phosphorylation Ser / Thr Kinases PKB / AKT
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Akt Kinase Activity Assay Kit (ab139436)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (1)References (8)

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Analysis of purified active Akt activity with ab139436.

    Key features and details

    • Assay type: Enzyme activity
    • Detection method: Colorimetric
    • Platform: Microplate reader
    • Sample type: Adherent cells, Purified protein, Suspension cells, Tissue Extracts

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    Overview

    • Product name

      Akt Kinase Activity Assay Kit
      See all AKT kits
    • Detection method

      Colorimetric
    • Precision

      Intra-assay
      Sample n Mean SD CV%
      Sample < 10%
      Inter-assay
      Sample n Mean SD CV%
      Sample < 10%
    • Sample type

      Adherent cells, Suspension cells, Tissue Extracts, Purified protein
    • Assay type

      Enzyme activity
    • Product overview

      Akt Kinase Activity Assay Kit (ab139436) is a non-radioactive assay providing a safe, rapid and reliable method for the screening of inhibitors or activators of Akt and for quantitating the activity of Akt in purified or partially purified enzyme preparations. This kit is based on a solid phase enzyme-linked immuno-absorbent assay (ELISA) that utilizes a specific synthetic peptide as a substrate for Akt and a polyclonal antibody that recognizes the phosphorylated form of the substrate. The assay is designed for the analysis of Akt activity in the solution phase. For the measurement of Akt in partially purified and purified enzyme preparations from any species.


      The kit offers the following advantages:


      1. Safe - non-radioactive measurement of kinase activity


      2. Flexible - kinetic and end-point options available


      3. Fast - results in < 4.5 hours


      4. Efficient - only 30 µl diluted sample needed per well

    • Platform

      Microplate reader

    Properties

    • Storage instructions

      Please refer to protocols.
    • Components 1 x 96 tests
      20X Wash Buffer 1 x 30ml
      Active Akt 1 x 30µl
      Akt Phosphospecific Substrate Antibody 1 x 5ml
      Akt Substrate Microtiter Plate 1 unit
      Antibody Dilution Buffer 1 x 10ml
      Anti-Rabbit IgG: HRP Conjugate 1 x 20µl
      ATP 1 x 2mg
      Kinase Assay Dilution Buffer 1 x 10ml
      Stop Solution 2 1 x 10ml
      TMB Substrate 1 x 10ml
    • Research areas

      • Signal Transduction
      • Protein Phosphorylation
      • Ser / Thr Kinases
      • PKB / AKT
      • Cancer
      • Signal transduction
      • Protein phosphorylation
      • Serine/threonine kinases
      • AKT
      • Kits/ Lysates/ Other
      • Kits
      • Cell Signaling Kits
      • Kinase kits
    • Relevance

      AKT, also known as protein kinase B (PKB), is a serine/threonine protein kinase. There are three mammalian isoforms of AKT: AKT1 (PKB alpha), AKT2 (PKB beta) and AKT3 (PKB gamma) with AKT2 and AKT3 being approximately 82% identical with the AKT1 isoform. Each isoform has a pleckstrin homology (PH) domain, a kinase domain and a carboxy terminal regulatory domain. AKT was originally cloned from the retrovirus AKT8, and is a key regulator of many signal transduction pathways. Its tight control over cell proliferation and cell viability are manifold; overexpression or inappropriate activation of AKT has been seen in many types of cancer. AKT mediates many of the downstream events of phosphatidylinositol 3 kinase (a lipid kinase activated by growth factors, cytokines and insulin). PI3 kinase recruits AKT to the membrane, where it is activated by PDK1 phosphorylation. Once phosphorylated, AKT dissociates from the membrane and phosphorylates targets in the cytoplasm and the cell nucleus. AKT has two main roles: (i) inhibition of apoptosis; (ii) promotion of proliferation. AKT has been shown to play a role in such metabolic processes as glucose transport, glycogen synthesis, glycolysis, and protein synthesis. It had also been shown to promote cell survival by inhibiting apoptosis through its ability to phosphoylate and inactivate several targets, including Bad, Forkhead transcription factors, and caspase 9. Activity of AKT has been associated with the phosphorylation of two sites: T308, in the activation loop of the kinase, and S473, at the carboxyl terminus. Phosphorylation of both sites contributes to AKT activity, however phosphorylation of T308 has been shown to be absolutely essential for AKT activation.
    • Cellular localization

      Cell Membrane, Cytoplasmic and Nuclear. Note=Nucleus after activation by integrin-linked protein kinase 1 (ILK1).
    • Alternative names

      • AKT1
      • C AKT
      • PKB
      • PRKBA
      • RAC
      • RAC alpha
      • RAC alpha serine/threonine-protein kinase
      • RAC PK alpha
      • v akt murine thymoma viral oncogene homolog 1
      see all

    Associated products

    • Related Products

      • SC-66, Allosteric Akt inhibitor (ab120802)
      • Triciribine, inhibitor of Akt activation (ab120936)
      • (-)-Deguelin, Akt inhibitor (ab120941)
      • Isobavachalcone, Akt inhibitor (ab141168)
      • SH-5, Akt inhibitor (ab141442)
      • Perifosine, Akt inhibitor (ab142055)
      • Hexamethylene bisacetamide (HMBA), cell differentiation inducer (ab142438)

    Images

    • Analysis of purified active Akt activity with ab139436.
      Analysis of purified active Akt activity with ab139436.

      Varying quantities of purified active Akt were assayed using the Akt substrate microtiter plate. Assays were incubated for 60 minutes at 30°C.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (8)

    Publishing research using ab139436? Please let us know so that we can cite the reference in this datasheet.

    ab139436 has been referenced in 8 publications.

    • Tsoi H  et al. Overexpression of BQ323636.1 Modulated AR/IL-8/CXCR1 Axis to Confer Tamoxifen Resistance in ER-Positive Breast Cancer. Life (Basel) 12:N/A (2022). PubMed: 35054486
    • Abdelnaby RM  et al. Dual PI3K/Akt Inhibitors Bearing Coumarin-Thiazolidine Pharmacophores as Potential Apoptosis Inducers in MCF-7 Cells. Pharmaceuticals (Basel) 15:N/A (2022). PubMed: 35455425
    • Leo MD  et al. TMEM16A channel upregulation in arterial smooth muscle cells produces vasoconstriction during diabetes. Am J Physiol Heart Circ Physiol 320:H1089-H1101 (2021). PubMed: 33449847
    • Kitchen P  et al. Targeting Aquaporin-4 Subcellular Localization to Treat Central Nervous System Edema. Cell 181:784-799.e19 (2020). PubMed: 32413299
    • Chen YH  et al. Mitochondrial Akt Signaling Modulated Reprogramming of Somatic Cells. Sci Rep 9:9919 (2019). PubMed: 31289326
    • Kosalai ST  et al. EZH2 upregulates the PI3K/AKT pathway through IGF1R and MYC in clinically aggressive chronic lymphocytic leukaemia. Epigenetics 14:1125-1140 (2019). PubMed: 31216925
    • Miller WP  et al. Deletion of the Akt/mTORC1 Repressor REDD1 Prevents Visual Dysfunction in a Rodent Model of Type 1 Diabetes. Diabetes 67:110-119 (2018). PubMed: 29074598
    • Bai D  et al. Hyperglycemia and hyperlipidemia blunts the Insulin-Inpp5f negative feedback loop in the diabetic heart. Sci Rep 6:22068 (2016). Functional Studies . PubMed: 26908121

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Question

    Akt Kinase Activity Assay Kit, ab139436 - which AKT activity is detected? All AKT (1,2 and 3) or just AKT1 activity?

    Read More

    Abcam community

    Verified customer

    Asked on Sep 30 2015

    Answer

    The AKT activity assay ab139436 measures activity of all three AKT, 1, 2, and 3.

    Read More

    Abcam Scientific Support

    Answered on Sep 30 2015

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