| Assay Method Information | |
| | LRRK2 Kinase Activity Assay |
| Description: | LRRK2 Kinase reactions were carried out in 384-well white polystyrene plates in a final volume of 6 μl using ADP-Glo Kinase Assay kit (Promega Corp.). Compound and substrates (LRRKtide peptide and ATP) in assay buffer were first dispensed in wells. Kinase reaction was then started by the addition of human recombinant LRRK2 protein. After 1h-incubation at 37 C., the enzymatic reaction was stopped by the addition of 6 μl of ADP-Glo Reagent-1 and an additional 40-minutes incubation at 23 C. (residual ATP depletion). A final 30-minutes incubation after 12 μL reagent-2 addition (ADP to ATP conversion and luciferin/luciferase reaction) was performed before luminescent signal acquisition (EnVision multimode plate reader-PerkinElmer, Inc.). Data from 10 individual concentrations of tested compounds (N=2) were fitted (XLfit -ID Business Solutions Ltd) to deliver IC50s (compound concentration leading to 50% inhibition of reference enzymatic activity). |
| Affinity data for this assay | |
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