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Our scientists can help you choose the right assay format, substrate, and enzyme for your experiment.
About AKT1
AKT1 (Protein Kinase B alpha) is a serine/threonine kinase and the founding member of the Akt family, which also includes AKT2 and AKT3. Activated downstream of PI3K upon receptor tyrosine kinase or GPCR stimulation, AKT1 requires phosphorylation at Thr308 by PDK1 and Ser473 by mTORC2 for full activation. Once active, AKT1 phosphorylates a broad array of substrates including GSK-3alpha/beta, FOXO transcription factors, TSC2, MDM2, BAD, and PRAS40, coordinating cell survival, proliferation, metabolism, and protein synthesis. AKT1 sits at the nexus of the PI3K/AKT/mTOR pathway, one of the most frequently dysregulated axes in human cancer. Activating mutations—most notably the E17K hotspot mutation—amplification of the AKT1 gene, and upstream loss of PTEN drive oncogenic signaling across breast, colorectal, ovarian, and endometrial cancers. AKT1 is a high-priority therapeutic target, with multiple isoform-selective and pan-AKT inhibitors in clinical development.
Why PhosphoSens for AKT1?
AKT1 drug discovery presents several assay challenges: the enzyme requires precise activation state management, physiological ATP concentrations are critical because many allosteric inhibitors show strong ATP-dependence, and distinguishing isoform-selective compounds from pan-AKT inhibitors demands sensitive kinetic discrimination. Endpoint assays such as ADP-Glo and HTRF capture only a single time point, missing inhibition mechanism nuances and risking false results from compound interference with luciferase or fluorescent antibodies. Radiometric assays add throughput and safety constraints. PhosphoSens continuous kinase assays monitor substrate phosphorylation in real time at physiological ATP, generating full progress curves that reveal allosteric versus ATP-competitive inhibition modes, enable accurate Ki determination, and detect time-dependent inhibition kinetics including covalent inactivation parameters kinact and KI—all in a homogeneous, no-antibody, mix-and-read format ideal for high-throughput screening and mechanistic profiling.
Key Pathways:
PI3K/AKT/mTOR signaling
PTEN tumor suppressor pathway
Insulin/IGF-1 receptor signaling
FOXO transcription factor regulation
Approved Inhibitors:
Capivasertib (Truqap, HR+/HER2- breast cancer in combination with fulvestrant)
Talk to a Scientist
Have questions about AKT1 assay design, selectivity panels, or covalent inhibitor characterization?
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How pricing works:
Select a kit or substrate below, then request a quote. Bundle discounts apply automatically.
Select a kit or substrate below, then request a quote. Bundle discounts apply automatically.
Real-Time AKT1 Activity Assays
Continuous, real-time fluorescent assays optimized for quantitative AKT1 activity measurements, IC50 determination, and mechanistic studies.
Assay Format: Continuous kinetic fluorescence (real-time)
PhosphoSens-Kinetic assays directly quantify enzyme activity by continuously monitoring substrate phosphorylation or dephosphorylation in real time, generating a full progress curve in every well.
Learn more about PhosphoSens-Kinetic →Need pricing or availability? Select a kit or substrate to request a quote below.
Kits
PhosphoSens-Kinetic Kinase Activity Kit - AQT0536
Ready-to-use assay kits containing substrate and all essential reagents.
Kit Contents
- Optimized PhosphoSens® substrate (AQT0536)
- ATP, DTT, EGTA, Reaction Buffer & Dilution Buffer
Automatically save 10% when bundling 10ug recombinant enzyme with your 1,000 assay kit. View enzymes
Price: $425
Substrate
PhosphoSens Kinase Substrate AQT0536
Bulk PhosphoSens® substrate for assay development and high-throughput workflows.
Specifications
Automatically save 10% when bundling Reagent Packs with your substrate. View Reagent Packs
Price: $2795
Ready to request pricing for AKT1?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteTRF AKT1 Screening Assays
Robust endpoint TRF assays optimized for high-throughput screening and quantitative AKT1 activity measurement in plate-based workflows.
Assay Format: Time-resolved fluorescence (TRF, endpoint)
PhosphoSens-Red assays use europium-based time-resolved fluorescence detection to enable robust, plate-based quantification of enzyme activity, making them well suited for high-throughput screening workflows.
Learn more about PhosphoSens-Red →Need pricing or availability? Select a kit or substrate to request a quote below.
Kits
PhosphoSens-Red Kinase Activity Kit - AQT0569
Ready-to-use TRF assay kits containing substrate and essential reagents.
Kit Contents
- Optimized PhosphoSens® substrate (AQT0569)
- ATP, DTT, EGTA, Reaction Buffer & Dilution Buffer
- Europium Detection Solution
Automatically save 10% when bundling 10ug recombinant enzyme with your 1,000 assay kit. View enzymes
Price: $455
Substrate
PhosphoSens Kinase Substrate AQT0569
Bulk substrate for PhosphoSens-Red TRF assays and high-throughput workflows.
Specifications
Automatically save 10% when bundling Reagent Packs with your substrate. View Reagent Packs
Price: $2795
Ready to request pricing for AKT1?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteActive Recombinant Enzymes for AKT1
Wild-type and mutant forms of AKT1 for assay development, kinase profiling, and mechanistic studies. Enzymes are supplied active and optimized for PhosphoSens® substrates.
AKT1, Active Full Length
Human • Baculovirus-Insect Cells
Technical Specifications
- Catalog: AQT-A16-10G
- Mutation: Full Length
- Species: Human
- Expression System: Baculovirus-Insect Cells
- Storage Buffer: Supplied as sterile 50 mM Tris-HCl, pH 7.5, 50-300mM NaCl, 10mM glutathione, 0.1mM EDTA, 0.25-1.0mM DTT, 0-0.1mM PMSF, 10-25% glycerol
- Molecular Weight: ~85 kDa
- Purity/Activity: For specific information on a given lot, see related technical data sheet.
- Storage Conditions: Store product at -70°C. For optimal storage, aliquot target into smaller quantities after centrifugation and store at recommended temperature. For most favorable performance, avoid repeated handling and multiple freeze/thaw cycles.
Available Sizes:
Price: $$278.00
Request a QuoteFrequently Asked Questions: AKT1 Assays
What ATP concentration does the AssayQuant AKT1 assay use, and why does it matter?
The PhosphoSens AKT1 assay is validated at physiological ATP concentrations (typically 1 mM), which is critical because many AKT inhibitorsu2014particularly allosteric compounds like MK-2206u2014display pronounced ATP-dependence in their potency. Using sub-physiological ATP, as required by ADP-Glo or coupled enzyme assays, can artificially inflate the apparent potency of ATP-competitive inhibitors and obscure the selectivity profile of allosteric candidates. Running assays at physiological ATP ensures IC50 and Ki values translate reliably to cellular and in vivo settings.
Can the PhosphoSens AKT1 assay distinguish allosteric inhibitors from ATP-competitive inhibitors?
Yes. Because PhosphoSens generates continuous progress curves, researchers can apply kinetic modeling to differentiate competitive, non-competitive, and allosteric inhibition modes in a single experiment. Allosteric inhibitors such as MK-2206 produce characteristic progress curve signatures distinct from ATP-competitive compounds, allowing mechanism-of-action classification without secondary assays. This capability is unavailable with endpoint formats like HTRF or ADP-Glo, which provide only a single inhibition value per condition.
Is the AssayQuant AKT1 assay suitable for measuring covalent or irreversible inhibitors?
Absolutely. The real-time nature of PhosphoSens assays makes them ideally suited for characterizing covalent inhibitors by directly measuring time-dependent loss of AKT1 activity and enabling kinact/KI determination from progress curve data. This is a significant advantage over endpoint assays, which cannot distinguish reversible from irreversible inhibition without extensive additional experiments. Researchers developing covalent AKT inhibitor scaffolds can obtain complete mechanistic parameters in a single continuous fluorescence read.
Data & Resources
Explore data and documents to support your kinase and phosphatase experiments. Download sample data, protocols and other resources to see how our assays perform and to help you get started in your own lab. All validation data generated using PhosphoSens® assays under recommended conditions.
Sample Report: AQT0734 - EGFR Assay Validation Report
Each validation report provides experimental conditions and data showing:
- Enzyme titration and linearity
- Sensor peptide substrate titration and Km determination
- ATP titration and Km determination
- DMSO titration and assay tolerance
- Inhibitor titration and IC50 determination
Protocol
Inhibitor IC50 Protocol
See how the PhosphoSens-Kinetic Assay can be used to find the IC50 of a kinase inhibitor.
Download Protocol →Webinar
Webinar: Continuous Assay Validation
Discover how continuous assay formats power deep understanding of kinase function. See how PhosphoSens® assays guide inhibitor profiling, selectivity assessment, and mechanistic characterization.
Watch the Webinar →Service
KinSight™ AKT1 Profiling
Need broader selectivity data? KinSight profiling runs your compounds across our full kinase panel under identical PhosphoSens conditions.
Learn about KinSight →Stay Informed
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