Assay Report Card

Basic Information

ID: ALA3888346

Type: Binding

Description: Kinase Activity Assay: The kinase reaction buffer consists of 50 mM Tris pH 8.5, 0.01% BRIJ-35, 10 mM MgCl2, 1 mM EGTA, and 2 mM DTT. In the assay, 1 nM LRRK2 WT or 250 pM LRRK2 G2019S kinase is incubated with the test compound (typically at 0 to 30 μM) for 30 minutes before the kinase reaction is initiated by addition of 1.3 mM ATP and 0.4 μM fluorescein-LRRKtide. The reaction mixture (20 μl total volume) is incubated for 2 hours at 30 °C. before the reaction is terminated by addition of 10 mM EDTA and 1 nM terbium-labelled anti-phospho-LRRKtide antibody (final volume 20 μl). The mixture is further incubated for 30 minutes at RT. TR-FRET is measured by excitation of the terbium-donor with 340 nm light and subsequent (delay time 100 μs) measurement of terbium and fluorescein emission at 495 nm and 520 nm, respectively, over a time window of 1000 μs. The measurement is repeated 10 times for fluorescein and 10 times for terbium emission with a 2000 μs time window between repeats. TR-FRET measurements are performed on a Biomek Synergy plate. The TR-FRET signal is calculated as the emission-ratio at 520 nm over 495 nm.

Activity Charts

Compound Summary

Parent Molecular WeightALogPPolar Surface Area
392.462.642.64
412.933.663.66
396.473.143.14
377.453.093.09
426.913.303.30
378.483.003.00
377.453.093.09
364.453.013.01
334.433.163.16
426.913.303.30
376.462.842.84
333.403.253.25
392.462.642.64
422.492.652.65
392.513.313.31
332.413.663.66
335.422.562.56
351.412.862.86
365.443.203.20
408.513.013.01
349.443.503.50
412.933.663.66
317.353.743.74
406.492.572.57
378.483.003.00
406.492.952.95
392.513.043.04
410.452.782.78
396.473.143.14