Assay Method Information
|Fluorescence Polarization Assay
|For competitive binding assays, 1 mL of 200 mM EphA4-LBD was pre-incubated with the tested compounds at various concentrations in 98 mL of PBS (pH 7.2) in 96-well black plates at roomtemperature for 10 min, after which 1 mL of 500 mMFITC-labeled EphA4 peptide was added to produce a final volume of 100 mL. KYL and DMSO were incubated in each assay as positive and negative controls, respectively. After 30 min of incubation at room temperature, the polarization values in millipolarization units were measured at excitation/emission wavelengths of 480/535 nm with a multilabel plate reader (PerkinElmer).
|Affinity data for this assay
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