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Our scientists can help you choose the right assay format, substrate, and enzyme for your experiment.
About WEE1
WEE1 (WEE1 G2 Checkpoint Kinase) belongs to the WEE family of nuclear tyrosine kinases and serves as a critical guardian of genomic integrity. WEE1 phosphorylates CDK1 at Tyr15 and CDK2 at Tyr15, inactivating these cyclin-dependent kinases to enforce the G2/M and S-phase checkpoints, respectively. This phosphorylation prevents mitotic entry until DNA repair is complete. WEE1 activity is itself regulated by ATR/ATM-mediated checkpoint signaling, PLK1-dependent degradation, and autophosphorylation. In cancer, WEE1 is frequently overexpressed, allowing tumor cells to survive replication stress by sustaining checkpoint arrest rather than undergoing apoptosis. Conversely, p53-deficient tumors rely heavily on the WEE1-CDK1 axis as their primary DNA damage checkpoint, rendering them exquisitely sensitive to WEE1 inhibition. This synthetic lethality rationale has driven intense drug discovery interest, particularly in combination with DNA-damaging agents, PARP inhibitors, and ATR inhibitors across ovarian, uterine, and hematologic malignancies.
Why PhosphoSens for WEE1?
WEE1 presents distinct assay challenges: its dual-specificity kinase activity on CDK1/CDK2 tyrosine substrates demands carefully optimized peptide substrates that reflect physiological context, and its relatively low intrinsic catalytic rate can make endpoint assays prone to signal artifacts at low ATP concentrations. ADP-Glo and HTRF approaches require fixed time-point sampling, risking nonlinear kinetics and missing early inhibitor-driven rate changes critical for covalent or slow-binding compounds. Radiometric assays add throughput and safety burdens. PhosphoSens continuous fluorescent kinase assays resolve these issues by generating real-time progress curves at physiological ATP concentrations, enabling direct KM(ATP) determination, accurate IC50 measurements, and full kinact/KI characterization for irreversible inhibitors — all without antibodies or coupled enzymes that can confound WEE1's moderate activity profile.
Key Pathways:
G2/M DNA Damage Checkpoint
DNA Replication Stress Response
CDK1/Cyclin B Regulation
ATR-CHK1 Signaling
Approved Inhibitors:
Adavosertib (AZD1775) - not FDA-approved; None approved
Talk to a Scientist
Have questions about WEE1 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 WEE1 Activity Assays
Continuous, real-time fluorescent assays optimized for quantitative WEE1 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 - AQT0728
Ready-to-use assay kits containing substrate and all essential reagents.
Kit Contents
- Optimized PhosphoSens® substrate (AQT0728)
- 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 AQT0728
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 WEE1?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteTRF WEE1 Screening Assays
Robust endpoint TRF assays optimized for high-throughput screening and quantitative WEE1 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 - AQT0728
Ready-to-use TRF assay kits containing substrate and essential reagents.
Kit Contents
- Optimized PhosphoSens® substrate (AQT0728)
- 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 AQT0728
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 WEE1?
Select a kit, substrate, or enzyme above. Our team will confirm pricing, availability, and any applicable bundle discounts.
Request a QuoteActive Recombinant Enzymes for WEE1
Wild-type and mutant forms of WEE1 for assay development, kinase profiling, and mechanistic studies. Enzymes are supplied active and optimized for PhosphoSens® substrates.
WEE1, Active Full Length
Human • Baculovirus-Insect Cells
Technical Specifications
- Catalog: AQT-W01-11G
- 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: MW 115 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: WEE1 Assays
What substrate does AssayQuant recommend for WEE1 kinase assays?
AssayQuant offers PhosphoSens peptide substrates derived from the CDK1 Tyr15 phosphorylation site, optimized for WEE1 catalytic activity. These substrates incorporate the proprietary Sox-based fluorophore to provide continuous, real-time signal output proportional to phosphorylation state. Substrate selection is validated against WEE1 kinetic parameters to ensure linear assay windows suitable for inhibitor profiling.
Can PhosphoSens WEE1 assays be used to characterize covalent inhibitors?
Yes u2014 PhosphoSens continuous kinetics are ideally suited for covalent inhibitor characterization because real-time progress curves directly capture the time-dependent loss of enzyme activity needed to calculate kinact and KI. Unlike endpoint assays that provide only a snapshot, full progress curve analysis with PhosphoSens accurately resolves slow-binding and irreversible inhibition mechanisms critical for next-generation WEE1 inhibitor development.
Why is physiological ATP concentration important for WEE1 inhibitor IC50 measurements?
Many WEE1 inhibitors are ATP-competitive, so IC50 values are highly sensitive to ATP concentration used in the assay. PhosphoSens assays run at or near physiological ATP levels (1-5 mM), producing IC50 data that is more predictive of cellular potency compared to the artificially low ATP concentrations (typically 10 u00b5M) required by luminescence-based ADP-detection platforms. This reduces the need for Cheng-Prusoff corrections and improves translation to cellular and in vivo models.
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™ WEE1 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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