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Our scientists can help you choose the right assay format, substrate, and enzyme for your experiment.
About AKT3
AKT3 (AKT Serine/Threonine Kinase 3, also known as PKBγ) belongs to the AGC kinase superfamily alongside AKT1 and AKT2. All three isoforms share a conserved domain architecture comprising a pleckstrin homology (PH) domain, a kinase domain, and a hydrophobic motif. AKT3 is activated downstream of PI3K via phosphoinositide-dependent kinase-1 (PDK1)-mediated phosphorylation at T305 and mTORC2-mediated phosphorylation at S472. Once active, AKT3 phosphorylates substrates including GSK3β, FOXO transcription factors, TSC2, and BAD, driving cell survival, proliferation, and metabolic reprogramming. AKT3 is predominantly expressed in the brain and is the dominant isoform in melanoma and glioblastoma. PTEN loss, PIK3CA gain-of-function mutations, and AKT3 amplification are frequent oncogenic drivers. The isoform-selective nature of AKT3 in CNS malignancies makes it an attractive target for precision oncology strategies, and understanding isoform-specific kinetics is critical for drug candidate differentiation.
Why PhosphoSens for AKT3?
A principal challenge in AKT3 drug discovery is distinguishing isoform-selective inhibition from pan-AKT activity in biochemical assays. Traditional endpoint methods such as ADP-Glo and HTRF measure single time-point signals that obscure mechanistic details and are highly sensitive to assay timing artifacts and compound interference. Radiometric assays offer sensitivity but introduce radioactive handling burdens and poor throughput. PhosphoSens continuous kinase assays overcome these limitations by generating real-time fluorescent progress curves using a sox-based peptide substrate at physiological ATP concentrations. This enables accurate Km, Vmax, and Ki determination, direct measurement of covalent kinact/KI parameters for irreversible or covalent-warhead inhibitors, and detection of slow-binding kinetics often missed at endpoint. The continuous format is fully compatible with AKT3 isoform selectivity profiling, reducing false positives and delivering mechanistically richer SAR data earlier in the discovery pipeline.
Key Pathways:
PI3K/AKT/mTOR signaling
PTEN tumor suppressor pathway
FOXO transcription factor regulation
GSK3 signaling
Approved Inhibitors:
Capivasertib (Truqap, HR+/HER2- breast cancer in combination with fulvestrant)
Talk to a Scientist
Have questions about AKT3 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 AKT3 Activity Assays
Continuous, real-time fluorescent assays optimized for quantitative AKT3 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 AKT3?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteTRF AKT3 Screening Assays
Robust endpoint TRF assays optimized for high-throughput screening and quantitative AKT3 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 AKT3?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteActive Recombinant Enzymes for AKT3
Wild-type and mutant forms of AKT3 for assay development, kinase profiling, and mechanistic studies. Enzymes are supplied active and optimized for PhosphoSens® substrates.
AKT3, Active Full Length
Human • Baculovirus-Insect Cells
Technical Specifications
- Catalog: AQT-A18-10G
- 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: $$348.00
Request a QuoteFrequently Asked Questions: AKT3 Assays
How does PhosphoSens enable AKT3 isoform selectivity profiling?
PhosphoSens uses validated peptide substrates optimized for AKT family members, allowing side-by-side kinetic profiling of AKT1, AKT2, and AKT3 under identical, physiological ATP conditions. Continuous progress curves reveal differences in inhibitor potency, residence time, and binding kinetics across isoforms that single-endpoint assays routinely miss. This mechanistic resolution is essential for advancing isoform-selective candidates with the right selectivity profile.
Can PhosphoSens AKT3 assays detect covalent or allosteric inhibitors?
Yes. The real-time continuous format is uniquely suited to characterize both covalent inhibitors, via kinact/KI determination from progress curve analysis, and allosteric inhibitors such as PH-domain binders, which can display slow-on or slow-off kinetics invisible to endpoint assays. This makes PhosphoSens particularly valuable for emerging AKT3 drug modalities beyond classical ATP-competitive inhibitors.
What are common assay artifacts when running AKT3 biochemical assays, and how are they avoided?
Inner filter effects, compound autofluorescence, and assay-timing variability at fixed endpoints can all produce false positives or negatives in conventional AKT3 assays. PhosphoSens continuous assays monitor the full fluorescent progress curve, allowing real-time identification of signal artifacts versus genuine inhibition. Running assays at physiological ATP concentrations also ensures inhibitor IC50 values are clinically translatable rather than artificially depressed by low-ATP conditions.
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™ AKT3 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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