What Differentiates PhosphoSens Substrates From Other Substrates?
PhosphoSens® Substrates are peptide substrates with our readout molecule, Sulfonamido-oxine (Sox), covalently attached in close proximity to the phosphorylation site.
As the enzyme acts on the substrate, phosphorylation induces a chelation‑enhanced fluorescence (ChEF) signal proportional to phosphorylation level.
This enables real-time, continuous measurement of phosphorylation.
Unlike traditional substrates used in endpoint assays (e.g., ELISA, radioactive, or mobility-shift assays), PhosphoSens substrates generate a direct fluorescent signal only when phosphorylated, eliminating the need for coupled reactions, secondary antibodies, or wash steps.
Key differentiators:
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Direct detection of phosphorylation: No downstream coupling enzymes or antibodies required.
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Continuous, real-time signal: Produces a true progress curve in every well, revealing kinetic parameters and inhibitor mechanisms.
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High sensitivity and specificity: Fluorescence is strictly dependent on enzymatic phosphorylation at a defined site.
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Flexible format: Compatible with purified enzymes, lysates, and physiological ATP concentrations.
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Streamlined workflow: Add-and-read, homogeneous format simplifies protocols and reduces variability.
Unlock the potential of your kinase research with AssayQuant's Built-For-Purpose Kinase Sensor Peptide Substrates
What Separates the PhosphoSens Workflow From Other Kinase Assay Technologies?
Understanding kinase and phosphatase inhibition is faster and easier with PhosphoSens.
Traditional assay platforms often rely on indirect measurements and are limited by the use of antibodies, coupled reactions, radioactivity, or complex instrumentation.
With a simple add-and-read protocol that generates a full progress curve in every well, you’ll get more data in less time with fewer wells.
What is The Significance of A Progress Curve in Every Well?
A continuous assay format that provides actionable data by revealing linearity or non-linearity in your progress curve, enables identification and characterization of progress curve lags, which are missed in an end-point format, and prevents you from reporting misleading information (such as an IC50 for an irreversible inhibitor) by accurately uncovering time dependent inhibition.
Empower Your Research and Drug Discovery With Kinetic Data Using AssayQuant's PhosphoSens Continuous Assay Format
How Can Reporting IC50 Values Be Misleading?
Not all kinase inhibitors are alike. In some cases, the compound’s degree of target inhibition varies with time—known as time-dependent inhibition (TDI).
A continuous assay, like PhosphoSens, measures the catalytic event of phosphorylation in real time enables determination of kinetic parameters that power your drug discovery research.
Watch our short video “The Role of Time-Dependent Inhibition Kinetics in Lead Discovery and Optimization” to learn more and download a copy of the presentation below.
What Are the Advantages of Kinome Profiling with a Continuous Assay?
There are many unique advantages of continuous assays over their endpoint counterparts. PhosphoSens reveals hidden insights within raw data, and generates progress curves that can help you identify essential aspects of an inhibitor's mode of action.
Uncover the power and kinetic insights of a continuous assay for kinome profiling with our newest webinar, available on demand!
Can PhosphoSens Measure Phosphatase Activity?
Protein phosphatases are an important drug target in the development of cancer therapies. With a renewed focus on development of these drug candidates, there is a need for robust assays that monitor phosphatase enzyme activity.
Explore our scientific poster to see how we harnessed the power of our PhosphoSens technology using to develop assays continuously monitor protein phosphatase activity.
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