Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Sample type: Adherent cells, Purified protein, Suspension cells, Tissue Extracts
Product nameAkt Kinase Activity Assay Kit
See all AKT kits
Intra-assay Sample n Mean SD CV% Sample < 10% Inter-assay Sample n Mean SD CV% Sample < 10%
Sample typeAdherent cells, Suspension cells, Tissue Extracts, Purified protein
Assay typeEnzyme activity
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
Storage instructionsPlease 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
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)
ab139436 has been referenced in 5 publications.
- 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