PRODUCT

Endpoint Time-Resolved Fluorescence (TRF) Assays

Screen even low signal kinases over a broad dynamic range in high-throughput

Red-shifted time resolved readout for primary screening applications

Based on proprietary technology developed at MIT, PhosphoSens-Red Assays deliver high-sensitivity measurements with time-resolved fluorescence (TRF), capturing both kinetic and time point data with exceptional signal stability. These assays enable real-time monitoring at the substrate level, providing in-depth insights into enzyme kinetics, inhibitor potency, and interaction dynamics, all while reducing background noise for superior data clarity.

Enhanced Sensitivity and Broad Dynamic Range

Utilize TRF to generate clear, high-quality data, reducing background interference for greater accuracy and sensitivity.

Direct Measurement of Enzyme Activity 

Measures enzyme activity at the substrate level, avoiding the complications of indirect assays that require additional steps.

Physiologically Relevant Conditions 

Use biologically relevant peptide substrate sequences in assays that are compatible with low to physiological [mM] concentrations of ATP.

Single-Step, Homogenous Workflow

Achieve fast and reproducible results with a homogenous, single-step workflow without compromising data quality.

Looking for a kinetic assay with a continuous readout?

PhosphoSens Catalog Product Packaging
PRODUCT CONFIGURATIONS

What's in the box?

PhosphoSens Substrates

Designed for higher throughput applications, each PhosphoSens Senor Peptide Substrate can be purchased as lyophilized 1mg aliquots. Each mg is sufficient for ~2,500 assays in a low‐volume 384‐well plate (10μM sensor peptide in 25μL reaction volume).

PhosphoSens Substrate Systems

Designed for higher throughput applications AND ease of use, each PhosphoSens Senor Peptide Substrate System pairs 1mg aliquots of substrate with the required reaction reagents needed to perform the assay (ATP, DTT, EGTA, Enzyme Reaction Buffer, and Enzyme Dilution Buffer). Each system is sufficient for ~2,500 assays in a low‐volume 384‐well plate (10μM sensor peptide in 25μL reaction volume).

PhosphoSens-Red Discovery Kits

Designed for streamlined technology assessment, each discovery kit includes sufficient volumes of substrate and reaction reagents to perform 200 reactions in a low‐volume 384‐well plate (10μM sensor peptide in 25μL reaction volume).
*Discounted technology evaluation kits are available for first time users and for new projects. Please inquire.

How it works

Because many compound libraries contain auto-fluorescent compounds, AssayQuant offers PhosphoSens Red assays that allow detection at longer (>600 nm) wavelength, with the same excitation maximum, and use time-resolved fluorescence (TRF). We recommend using a 100 µsecond delay between excitation and detection at the emission wavelength, with a 300 µsecond acquisition time, to avoid signal contamination from fluorescent materials in the reaction mixture, which have a very short (nanosecond) lifetime.

Once the reaction is run, a Europium ion (Eu3+) based reagent is added to each well, which displaces Mg++ in the chelation complex. Excitation of the Soc fluorophore sensitizes the europium luminescence, producing light with a 615 nm (red) maximum that can be measured using standard filters.

 

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Didn’t find what you were looking for?

We also offer custom assay development and kinase and phosphatase profiling services. Tell us about your specific project, and our experts can help you select the right PhosphoSens tool for your kinetic assay needs.

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