Table 4HTRF assay protocol for the EGFR enzymes in 1536-well plate format

StepParameterValueDescription
1Enzyme2 nlEGFR at 3 nM final
2Compound22 nlLibrary Compounds in 0.128 to 10 mM titration series or control
3Reagent1.5 μl10 µM ATP and 0.25 µM substrate final
4Incubation30 minAmbient
5Reagent3 μlantibody and XL-665
6Incubation30 minAmbient
7DetectorEx 320, Em 615,665EnVision plate reader

From: HTS for Identification of Inhibitors against the ERK Signaling Pathway using a Homogenous Cell-based Assay

Cover of Probe Reports from the NIH Molecular Libraries Program
Probe Reports from the NIH Molecular Libraries Program [Internet].

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