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PhosphoSens CDK2/CycE2 Protein Kinase Assays, Substrates & Recombinant Enzymes
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About CDK2/CycE2
Cyclin-dependent kinase 2 (CDK2) is a serine/threonine kinase belonging to the CDK family, which serves as master regulators of cell cycle progression. CDK2 is activated by binding to its regulatory partner Cyclin E2 (encoded by CCNE2), forming the CDK2/CycE2 complex that drives the G1-to-S phase transition. This complex phosphorylates key substrates including Rb family members (p107, p130), NPAT, and CDC6, licensing DNA replication origin firing and promoting S phase entry. CDK2/CycE2 activity is negatively regulated by CIP/KIP family inhibitors such as p21 and p27, and by the INK4 proteins acting indirectly through CDK4/6. Dysregulation of CDK2 through CCNE2 amplification, loss of p27, or Rb pathway disruption is prevalent in breast, ovarian, gastric, and lung cancers. CDK2 inhibition represents a compelling oncology strategy, particularly in tumors with CDK4/6 inhibitor resistance or CCNE1/2 amplification, making selective CDK2 inhibitors highly sought after in contemporary drug discovery programs.
Why PhosphoSens for CDK2/CycE2?
CDK2/CycE2 assays present significant challenges because the kinase requires precise co-expression of its cyclin partner to achieve physiological activation, and endpoint assays such as ADP-Glo or HTRF risk introducing artifacts from incubation time selection and compound interference at luciferase or fluorophore readout steps. Radiometric assays offer sensitivity but are low-throughput and generate radioactive waste. PhosphoSens continuous kinetics technology uses a chemosensor-embedded peptide substrate that generates a real-time fluorescent signal directly proportional to phosphorylation, enabling full progress curve capture under physiological ATP concentrations. This approach directly measures covalent inhibitor kinact/KI parameters, detects partial inhibitors and unusual kinetics invisible to endpoint methods, and eliminates inner-filter and signal-quenching artifacts common in homogeneous assays. The result is higher-quality SAR data, accurate IC50 and Ki determination, and robust selectivity profiling for CDK2-targeted drug discovery programs.
Key Pathways:
Cell Cycle G1/S Transition
Rb/E2F Signaling
DNA Damage Response
PI3K/AKT/mTOR Pathway
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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 CDK2/CycE2 Activity Assays
Continuous, real-time fluorescent assays optimized for quantitative CDK2/CycE2 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 - AQT0297
Ready-to-use assay kits containing substrate and all essential reagents.
Kit Contents
- Optimized PhosphoSens® substrate (AQT0297)
- 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 AQT0297
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 CDK2/CycE2?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteNo PhosphoSens-Red format is currently available for CDK2/CycE2.
Ready to request pricing for CDK2/CycE2?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteNo recombinant enzymes are currently available for CDK2/CycE2.
Frequently Asked Questions: CDK2/CycE2 Assays
What substrate does AssayQuant use for CDK2/CycE2 assays and why is it suitable?
AssayQuant employs a proprietary PhosphoSens peptide substrate engineered with a Sox-based chemosensor that undergoes a fluorescence change upon phosphorylation by CDK2. The substrate is derived from validated CDK2 consensus sequences, ensuring relevant biochemical engagement. This design enables continuous, real-time signal generation without antibodies or secondary detection reagents.
Can the PhosphoSens CDK2/CycE2 assay be used to characterize covalent inhibitors?
Yes. Because PhosphoSens captures full kinetic progress curves in real time, it is ideally suited for measuring covalent inhibitor kinetics, including the inactivation rate constant (kinact) and inhibitor binding constant (KI). These parameters cannot be reliably extracted from single-timepoint endpoint assays, making continuous kinetics a critical advantage for covalent CDK2 inhibitor programs.
How does the CDK2/CycE2 assay perform at physiological ATP concentrations?
PhosphoSens assays are validated at physiological ATP concentrations (approximately 1 mM), which is essential for accurate competitive inhibitor IC50 determination and for avoiding the artificially potent IC50 values generated under low-ATP conditions used in many ADP-Glo or HTRF formats. Running assays near cellular ATP levels produces inhibitor potency data that more faithfully predicts cellular activity.
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™ CDK2/CycE2 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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