Overview

  • Product nameAKT Activity Assay Kit
    See all AKT kits
  • Tests
    40 x 1 assay
  • Sample type
    Tissue Extracts, Cell Lysate
  • Assay typeEnzyme activity
  • Product overview

    Abcam's AKT Activity Assay Kit utilizes an Akt-specific antibody to immunoprecipitate Akt from cell lysate. Activity of the Akt is then determined in a kinase reaction using recombinant GSK-3α as substrate. Phosphorylation of the GSK-3α can be analyzed by Western blot analysis using the phospho-GSK-3α specific antibody included in the kit. The kit specifically detects Akt1, Akt2, and Akt3 activities, other kinase activities would not be detected.
    Visit our FAQs page for tips and troubleshooting.

     

    The AKT Specific Antibody reacts with human, mouse, and rat.

  • Notes

    Akt is a protein kinase that can be activated by insulin and various growth factors and functions in a pathway involving PI3 kinase. Recent evidence suggests that Akt functions to promote cell survival by actively inhibiting apoptosis.

  • Tested applicationsFunctional Studiesmore details

Properties

  • RelevanceAKT, 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 localizationCell 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

Applications

Our Abpromise guarantee covers the use of ab65786 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
Functional Studies Use at an assay dependent dilution.

AKT Activity Assay Kit images

  • Western blot analysis of phospho-GSK-3 alpha in AKT negative (Lane 1) and AKT positive (Lane 2,3) Jurkat cell lysates using AKT Activity Assay Kit (ab65786).

Protocols

References for AKT Activity Assay Kit (ab65786)

This product has been referenced in:
  • Damera G  et al. An RGS4-mediated phenotypic switch of bronchial smooth muscle cells promotes fixed airway obstruction in asthma. PLoS One 7:e28504 (2012). Functional Studies . Read more (PubMed: 22253691) »
  • Miyazaki M  et al. Early activation of mTORC1 signalling in response to mechanical overload is independent of phosphoinositide 3-kinase/Akt signalling. J Physiol 589:1831-46 (2011). Read more (PubMed: 21300751) »

See all 3 Publications for this product

Product Wall

The kinase buffer contains PMSF, Na-vanadate and Na-pyrophosphate and EDTA. Typically nothing more is required to protect the Akt enzyme activity. Additional protease inhibitors might be added if desired. This should not affect the assay, but addition ...

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I have been informed by the lab that the antibody used in this kit is not a phospho specific Ab. The immunogen spans the N Terminal of the protein Akt.

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The AKT Activity Assay Kit (ab65786) can be used to detect akt activity in human, mouse, and rat tissue extracts and cell lysates.

I know the AKT can be detected in colon tumours, I don't know about liver....

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