EGFR Kinase Activity Solutions

Continuous, Direct, and Catalytic Protein Kinase Activity Assays

Product Information

The recommended PhosphoSens Substrate to assay EGFR activity in the PhosphoSens-Kinetic format is AQT0734

Optimized for EGFR: Study enzyme activity, generate kinetic data, discover inhibitors, characterize mechanisms of action (MOA), and more.

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Product Configurations

EGFR PhosphoSens-Kinetic Discovery Kit

Recommended Product: PhosphoSens-Kinetic Kinase Discovery Kit AQT0734

  • Designed for streamlined technology evaluation, PhosphoSens Discovery Kits enable 200 assays. Each kit includes substrate and all essential reaction reagents (ATP, DTT, EGTA, Enzyme Reaction Buffer and Enzyme Dilution Buffer), optimized for your convenience. Enzyme not included.

EGFR PhosphoSens-Kinetic Kinase Substrate

Recommended Product: PhosphoSens Kinase Substrate AQT0734

  • Flexible, scalable, and cost-effective! Available by the milligram, with each milligram enabling an average of 2,500 assays, PhosphoSens Substrates are ideal for high-throughput needs and custom workflows.

PhosphoSens-Kinetic Kinase Substrate System AQT0734

Recommended Product: PhosphoSens-Kinetic Kinase Substrate System AQT0734

  • Comprehensive and adaptable! These systems pair bulk substrate with sufficient supporting reagents (ATP, DTT, EGTA, Enzyme Reaction Buffer and Enzyme Dilution Buffer) to perform the full number of assays for each milligram of substrate purchased—giving you everything you need for seamless assay execution.
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How It Works

PhosphoSens continuous assays utilize a simple add-and-read protocol, providing the capability for real-time, kinetic measurements, with a straight-forward, fluorescence intensity (FI) measurement.
  • These assays provide high accuracy and precision by allowing measurement of the initial reaction rate (measured as the slope of the initial linear region), typically during the first 30-60 minutes and include many data points, with readings performed every 30 seconds to two minutes.
  • This continuous assay format also allows for correction of compound autofluorescence, because this background signal does not change over time and can be corrected for by subtracting background signals (includes compound and all components of the reaction besides the enzyme) from the total fluorescence (all components including enzyme) to determine the net signal at each time point.
  • The continuous format is compatible with essentially any commercially available fluorescence microplate reader capable of kinetic readings, and associated laboratory automation equipment. No stop solution or quench/develop step is needed, minimizing time, while maximizing throughput.
  • PhosphoSens-Kinetic Kinase Inhibitor IC50 Determination

    This protocol outlines assay conditions when working in a low‐volume 384‐well plate with 20 µL final well volume and 10 µM Sensor peptide at 1 mM ATP. It assumes that an optimal concentration of kinase and DMSO in the reaction has been previously determined or otherwise chosen. A 12-point compound titration (including a “0” compound concentration) should be run in duplicate using a 3fold dilution scheme for each compound with a separate blank at each compound concentration. Final assay concentrations will be the same as above except for the additional of 2% DMSO final with or without compound.
    VIEW PROTOCOL

    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
  • VIEW REPORT

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