PHOSPHOSENS® KINASE ASSAY CATALOG

Kinase Selectivity Profiling That Shows You More Than % Inhibition

KinSight™ profiles your compounds against 400+ validated kinases using continuous, real-time assays — delivering true selectivity data, TDI detection, and MOA insights that endpoint platforms systematically miss.

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THE PROBLEM WITH ENDPOINT PROFILING

Why Settle for a Single Snapshot of Enzyme Activity?

Most kinome profiling platforms take one measurement and call it done. But a single time point assumes the reaction is linear, stable, and uninhibited at that exact moment — an assumption that silently breaks down more often than you'd think.

The example below is real: four kinases that look nearly identical at endpoint. Progress curves tell a completely different story.

Linear Kinetics

Endpoint and kinetics fully agree.

Two clean straight lines from origin — DMSO rising steeply, compound shallow but linear. This is a straightforward reversible inhibitor, and the only pattern endpoint profiling reliably identifies correctly.

Initial Lag

Both curves flat ~20 min before rate establishes.

The enzyme takes time to reach its steady-state activity. Endpoint assays that sample during this lag calculate a falsely low rate, inflating apparent inhibition. Continuous assays identify the true linear window and calculate from there.

TDI Detected

Inhibition deepens over time — slope progressively decelerates.

Both curves start from a shared elevated baseline, then diverge as the compound progressively occupies or modifies the enzyme. A single endpoint measurement captures only a snapshot — and misses the mechanism and its true potency entirely.

Assay Artifact

Heavy scatter throughout — noise invisible at endpoint.

The compound curve shows extreme replicate variability. The 61% inhibition read is an artifact with no statistical basis. Endpoint platforms report this with the same confidence as a clean result.

Endpoint and kinetics fully agree.

Both curves flat ~20 min before rate establishes.

Inhibition deepens over time — slope progressively decelerates.

Heavy scatter throughout — noise invisible at endpoint.

The compound curve shows extreme replicate variability. The 61% inhibition read is an artifact with no statistical basis. Endpoint platforms report this with the same confidence as a clean result.

1. Submit your compound and select a panel

Choose from six predefined panels or work with our team to build a custom target list. Single concentration or dose-response available.

2. We run continuous assays at your ATP conditions

ATP Km for maximal potency insight, 1 mM for physiological relevance, or both. Every well generates a full progress curve — actual rates from multiple data points.

3. Expert review and annotation

Our scientists review every result and annotate TDI signals, unusual kinetics, and MOA insights. Not just a data dump — a report you can act on.

4. Full data package delivered in two weeks

Waterfall plots, kinome tree visualization, tabular % inhibition data, and annotated progress curves — ready to share with your team and advance your program.

Panel Name	Panel Size	ATP Options	Target List
Wild Type Protein Kinase Panel	401	ATP K_m and 1mM now offering custom ATP concentrations*	View Panel >
Full Protein Kinase Panel	450	ATP K_m and 1mM now offering custom ATP concentrations*	View Panel >
Tyrosine Kinase (TK) Panel	83	ATP K_m and 1mM now offering custom ATP concentrations*	View Panel >
Cyclin Dependent Kinase (CDK) Panel	32	ATP K_m and 1mM now offering custom ATP concentrations*	View Panel >
Mutant Protein Kinase Panel	53	ATP K_m and 1mM now offering custom ATP concentrations*	View Panel >
KiNode Panel	69	ATP K_m and 1mM now offering custom ATP concentrations*	View Panel >
Custom Panel	1-100	ATP K_m and 1mM now offering custom ATP concentrations*	Contact Us to Design >

*Kinase selectivity profiling is conducted on a bi-monthly schedule, with assays run at the apparent ATP Km (to enable accurate potency assessment) or at 1 mM ATP (to approximate physiological conditions). Alternative ATP concentrations can be accommodated upon request.

Platform comparison

Platform	Measures activity directly	Detects time dependent inhibition	Real-time kinetic readout	Detects all inhibitor types
AssayQuant PhosphoSens®
Luminescent assayse.g. Kinase-Glo, ADP-Glo
Radiometric assayse.g. ³³P/³²P filter binding
Mobility shift assayse.g. Caliper EZ Reader
Competition bindinge.g. KINOMEscan, KdELECT

WEBINAR

Deep Kinetic Characterization: When IC₅₀ Isn't Enough

A continuous assay, like our PhosphoSens assay, that measures the catalytic event of phosphorylation in real time enables determination of kinetic parameters that power your drug discovery research.

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BLOG

Continuous vs. Endpoint Kinase Assays: What You Need to Know

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PROTOCOL

PhosphoSens-Kinetic Phosphatase Inhibitor IC₅₀ Determination

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PhosphoSens® assays are continuous, real-time kinase activity assays that directly measure phosphorylation of a substrate peptide throughout the reaction. Unlike endpoint assays that capture a single time point, PhosphoSens generates a full progress curve — enabling true kinetic analysis including IC₅₀, Kᵢ, kobs, and time-dependent inhibition (TDI) from a single experiment.

PhosphoSens-Kinetic is a continuous fluorescence assay that monitors kinase activity in real time throughout the reaction, generating full progress curves. PhosphoSens-Red is a time-resolved fluorescence (TRF) endpoint format optimized for higher throughput screening. Both use the same underlying PhosphoSens® substrate technology — the choice depends on whether you need kinetic depth (Kinetic) or screening throughput (Red).

TDI compounds produce a characteristic change in the progress curve shape — the inhibition deepens over time as the compound slowly occupies or covalently modifies the enzyme. Because PhosphoSens monitors activity continuously, this curve deviation is directly visible. Endpoint assays that measure at a single time point will either miss TDI entirely or mischaracterize its potency, depending on when they sample the reaction.

PhosphoSens-Kinetic assays require a standard fluorescence plate reader capable of kinetic reads (repeated measurements over time) with excitation/emission appropriate for the Sox fluorophore. PhosphoSens-Red assays require a time-resolved fluorescence (TRF) reader. Most modern multimode readers in drug discovery labs are compatible. Contact us if you need compatibility confirmation for your specific instrument.

We offer custom assay development for kinase targets not currently in our catalog. Our team can design and validate a PhosphoSens substrate for your target, typically within 8–12 weeks. Contact us to discuss your target, timeline, and project requirements.

Ready to Characterize Your Compound?

Talk to one of our scientists about your target, your workflow, and which assay format will get you the data you need.

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Technology Overview

PhosphoSens-Kinetic

PhosphoSens-Lysate

PhosphoSens-Red

Proprietary Sox Sensors

Kinase Assays

Phosphatase Assays

Lysate Assays