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    akt-activity-assay-kit-ab65786.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases PKB / AKT
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AKT Activity Assay Kit (ab65786)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (1)Q&A (3)References (4)

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Functional Studies - AKT Activity Assay Kit (ab65786)

    Key features and details

    • Assay type: Enzyme activity
    • Sample type: Cell Lysate, Tissue Extracts

    You may also be interested in

    Protein
    Product image
    Recombinant human AKT1 protein (Active) (ab62279)

    View more associated products

    Overview

    • Product name

      AKT Activity Assay Kit
      See all AKT kits
    • Sample type

      Tissue Extracts, Cell Lysate
    • Assay type

      Enzyme 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

      This product is manufactured by BioVision, an Abcam company and was previously called K435 Akt Activity Assay Kit, KinaseSTAR. K435-40 is the same size as the 40 test size of ab65786.

      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.

    Properties

    • Storage instructions

      Store at -20°C. Please refer to protocols.
    • Components Identifier 40 tests
      Akt Specific Antibody Red 1 x 80µl
      GSK-3a Protein/ATP Mixture Blue 1 x 80µl
      Kinase Assay Buffer WM 1 x 25ml
      Kinase Extraction Buffer NM 1 x 80ml
      Phospho-GSK-3a Specific Antibody Green 1 x 50µl
      Protein A Sepharose Clear 1 x 2ml
    • Research areas

      • Signal Transduction
      • Protein Phosphorylation
      • Ser / Thr Kinases
      • PKB / AKT
      • Cancer
      • Signal transduction
      • Protein phosphorylation
      • Serine/threonine kinases
      • AKT
    • 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

    • Functional Studies - AKT Activity Assay Kit (ab65786)
      Functional Studies - AKT Activity Assay Kit (ab65786)
      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

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (4)

    Publishing research using ab65786? Please let us know so that we can cite the reference in this datasheet.

    ab65786 has been referenced in 4 publications.

    • He X  et al. Loss of hepatic aldolase B activates Akt and promotes hepatocellular carcinogenesis by destabilizing the Aldob/Akt/PP2A protein complex. PLoS Biol 18:e3000803 (2020). PubMed: 33275593
    • 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 . 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). PubMed: 21300751
    • Hussain AR  et al. Resveratrol Suppresses Constitutive Activation of AKT via Generation of ROS and Induces Apoptosis in Diffuse Large B Cell Lymphoma Cell Lines. PLoS One 6:e24703 (2011). PubMed: 21931821

    Customer reviews and Q&As

    Show All Reviews Q&A
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    1-4 of 4 Abreviews or Q&A

    Strong signal

    Excellent Excellent 5/5 (Ease of Use)
    Abreviews
    Abreviews
    abreview image
    Instructions for the use of the kit are clear.

    We used mouse cancer associated fibroblasts (CAF) treated with TGFb (described inductor of Akt signal) as positive control.

    The total-Akt antibody used for IP worked excellent and the pGSKa antibody for detection also.

    Results were consistents between the replica of the experiment.

    Abcam user community

    Verified customer

    Submitted Jun 18 2020

    Question

    1 - Does the kinase buffer contain protease inhibitors or phophotase inhibitors? If not, should they be added and how do they effect the experiment?
    2 - Are there any modifications for this protocol for tissue samples?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 17 2014

    Answer

    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 of more protease inhibitors and phosphatase inhibitors has not yet been tested.

    Tissue samples should be homogenized in the extraction buffer and then centrifuged to obtain a clarified homogenate which can be used in the assay.

    Read More

    Jeremy Kasanov

    Abcam Scientific Support

    Answered on Jan 17 2014

    Question

    Kindly contacted us to inquire about the Akt specific antibody used in this kit.

    Read More

    Abcam community

    Verified customer

    Asked on Jul 01 2013

    Answer

    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.

    Read More

    Keith B

    Abcam Scientific Support

    Answered on Jul 01 2013

    Question

    One more point. Do you know if your kit AKT Activity Assay Kit (ab65786) can be used to detect akt activity in liver samples,

    Read More

    Abcam community

    Verified customer

    Asked on Jul 10 2012

    Answer

    Thank you for contacting us.

    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.I would encourage youto consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about this specific research field.

    I hope this is helpful andI hope that the available literature in this area can provide some clarification about AKT in liver. Please do not hesitate to contact us again.

    Read More

    Abcam Scientific Support

    Answered on Jul 10 2012

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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