Key features and details
- Assay type: Enzyme activity
- Sample type: Cell Lysate, Tissue Extracts
Product nameAKT Activity Assay Kit
See all AKT kits
Sample typeTissue Extracts, Cell Lysate
Assay typeEnzyme activity
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-3a as substrate. Phosphorylation of the GSK-3a can be analyzed by Western blot analysis using the phospho-GSK-3a specific antibody included in the kit. The kit specifically detects Akt1, Akt2, and Akt3 activities, other kinase activities would not be detected.
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The AKT Specific Antibody reacts with human, mouse, and rat.
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.
Storage instructionsStore 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
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).
- C AKT
- 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)
ab65786 has been referenced in 3 publications.
- 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