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Assay Method Information |
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| | FRET-based Z'-lyte Assay |
| Description: | Briefly, 10 μL per well reactions contained ATP concentration at 25 μm for Syk,2 μm Tyr2 peptide substrate in 50 mm HEPES (PH 7.5), 0.01% BRIJ-35, 10 mm MgCl2, 1 mm EGTA, 0.0247 μg/mL Syk, and inhibitors as appropriate. The reaction wasperformed at room temperature for 1.5 h, and then, 5 μL of development reagent was added for a further 1-h room temperature incubation followed by the addition of 5 μL of stop solution. Fluorescence signal ratio of 460 nm/530 nm was examined on EnVision Multilabel Reader (Perkin-Elmer, Inc., Waltham, MA, USA). |
| Affinity data for this assay | |
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