| Assay Method Information | |
| | Kinase Assay (ALK) |
| Description: | Kinase Assay Protocol:Bar-coded Corning, low volume NBS, black 384-well plate (Corning Cat. #4514) 1. 2.5 uL-4x Test Compound or 100 nL 100x plus 2.4 uL kinase buffer; 2. 5 uL-2x Peptide/Kinase Mixture; 3. 2.5 uL-4xATP Solution; 4. 30-second plate shake; 5. 60-minute Kinase Reaction incubation at room temperature; 6. 5 uL-Development Reagent Solution; 7. 30-second plate shake; 8. 60-minute Development Reaction incubation at room temperature; 9. Read on fluorescence plate reader and analyze the data. Kinase-Specific Assay Conditions:The 2xALK/Tyr 01 mixture is prepared in 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MgCl2, 1 mM EGTA. The final 10 uL Kinase Reaction consists of 4.25-96 ng ALK and 2 uM Tyr 01 in 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MgCl2, 1 mM EGTA. After the 1 hour Kinase Reaction incubation, 5 uL of a 1:256 dilution of Development Reagent B is added. |
| Affinity data for this assay | |
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