#	Aladdin ID	Assay Type	Description	Organism	Compounds	Reference	BAO Format	Source
1.	ALA3889252	B	Homogeneous Time Resolved-fluorescence Resonance Energy Transfer (TR-FRET) Assay: LRRK2 kinase activity is measured using a LanthaScreen kinase activity assay available from Invitrogen (Life Technologies Corporation). The assay is a homogeneous time resolved-fluorescence resonance energy transfer (TR-FRET) assay that measures phosphorylation of a fluorescein-labelled peptide substrate (fluorescein-LRRKtide, fluorescein-GAGRLGRDKYKTLRQIRQ) (SEQ ID NO 1) as a result of LRRK2 kinase activity. The phosphorylated peptide is recognized by a terbium-labelled phospho-specific anti-LRRKtide antibody and, subsequently, the phosphorylated LRRKtide can be quantified by the extent of TR-FRET between the terbium donor and fluorescein acceptor.The LRRK2 kinase is obtained from Invitrogen (Life Technologies Corporation) and comprises residues 970 to 2527 of the full length human wild-type LRRK2 kinase or a similar sequence with the G2019S mutation. As discussed above, this mutation increases the kinase activity relative to the wild-type. The kinase reactions are performed in a 20 uL volume in 384-well plates. 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 uM) for 30 minutes before the kinase reaction is initiated by addition of 1.3 mM ATP and 0.4 uM fluorescein-LRRKtide. The reaction mixture (20 ul 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 ul). 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 us) measurement of terbium and fluorescein emission at 495 nm and 520 nm, respectively, over a time window of 1000 us. The measurement is repeated 10 times for fluorescein and 10 times for terbium emission with a 2000 us 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.		23		single protein format	BindingDB Database
2.	ALA3889253	B	Homogeneous Time Resolved-fluorescence Resonance Energy Transfer (TR-FRET) Assay: LRRK2 kinase activity is measured using a LanthaScreen kinase activity assay available from Invitrogen (Life Technologies Corporation). The assay is a homogeneous time resolved-fluorescence resonance energy transfer (TR-FRET) assay that measures phosphorylation of a fluorescein-labelled peptide substrate (fluorescein-LRRKtide, fluorescein-GAGRLGRDKYKTLRQIRQ) (SEQ ID NO 1) as a result of LRRK2 kinase activity. The phosphorylated peptide is recognized by a terbium-labelled phospho-specific anti-LRRKtide antibody and, subsequently, the phosphorylated LRRKtide can be quantified by the extent of TR-FRET between the terbium donor and fluorescein acceptor.The LRRK2 kinase is obtained from Invitrogen (Life Technologies Corporation) and comprises residues 970 to 2527 of the full length human wild-type LRRK2 kinase or a similar sequence with the G2019S mutation. As discussed above, this mutation increases the kinase activity relative to the wild-type. The kinase reactions are performed in a 20 uL volume in 384-well plates. 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 uM) for 30 minutes before the kinase reaction is initiated by addition of 1.3 mM ATP and 0.4 uM fluorescein-LRRKtide. The reaction mixture (20 ul 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 ul). 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 us) measurement of terbium and fluorescein emission at 495 nm and 520 nm, respectively, over a time window of 1000 us. The measurement is repeated 10 times for fluorescein and 10 times for terbium emission with a 2000 us 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.	Homo sapiens	23		single protein format	BindingDB Database
3.	ALA4031820	B	Inhibition of wild type recombinant human GST-tagged LRRK2 (970 to 2527 residues) expressed in baculovirus using fluorescein-LRRKtide as substrate after 2 hrs by TR-FRET assay	Homo sapiens	26		assay format	Scientific Literature
4.	ALA4031821	B	Inhibition of recombinant human GST-tagged LRRK2 G2019S mutant (970 to 2527 residues) expressed in baculovirus using fluorescein-LRRKtide as substrate after 2 hrs by TR-FRET assay	Homo sapiens	26		assay format	Scientific Literature
5.	ALA4031822	B	Inhibition of human wild type LRRK2 phosphorylation at ser935 transfected in HEK293 cells after 48 hrs by in-cell Western assay	Homo sapiens	1		cell-based format	Scientific Literature
6.	ALA4031823	B	Inhibition of human LRRK2 G2019S mutant phosphorylation at ser935 transfected in HEK293 cells after 48 hrs by in-cell Western assay	Homo sapiens	1		cell-based format	Scientific Literature
7.	ALA4031824	B	Inhibition of human LRRK2 A2016T mutant phosphorylation at ser935 transfected in HEK293 cells after 48 hrs by in-cell Western assay	Homo sapiens	1		cell-based format	Scientific Literature
8.	ALA4031825	B	Inhibition of human wild type LRRK2 phosphorylation at ser935 expressed in BacMam transduced HEK293 cells after 48 hrs by in-cell Western assay	Homo sapiens	13		cell-based format	Scientific Literature
9.	ALA4031826	B	Inhibition of human LRRK2 G2019S mutant phosphorylation at ser935 expressed in BacMam transduced HEK293 cells after 48 hrs by in-cell Western assay	Homo sapiens	13		cell-based format	Scientific Literature
10.	ALA4031827	B	Inhibition of human LRRK2 A2016T mutant phosphorylation at ser935 expressed in BacMam transduced HEK293 cells after 48 hrs by in-cell Western assay	Homo sapiens	1		cell-based format	Scientific Literature
11.	ALA4031828	B	Inhibition of human LRRK2 G2019S mutant phosphorylation at ser1292 expressed in BacMam transduced HEK293 cells after 48 hrs by in-cell Western assay	Homo sapiens	1		cell-based format	Scientific Literature
12.	ALA4031829	A	Apparent permeability across apical to basolateral side in MDCK cells expressing human MDR1	Canis lupus familiaris	13		cell-based format	Scientific Literature
13.	ALA4031830	A	Efflux ratio of apparent permeability in MDCK cells expressing human MDR1	Canis lupus familiaris	13		cell-based format	Scientific Literature
14.	ALA4031831	A	Clearance in rat liver microsomes	Rattus norvegicus	2		assay format	Scientific Literature
15.	ALA4031832	A	Clearance in human liver microsomes	Homo sapiens	13		assay format	Scientific Literature
16.	ALA4031833	F	In vivo inhibition of LRRK2 phosphorylation at ser935 in brain of C57BL/6 mouse at 25 mg/kg, po measured at 12 hrs post dose by Western blot analysis	Mus musculus	1		organism-based format	Scientific Literature
17.	ALA4031834	A	Unbound drug level in C57BL/6 mouse brain at 25 mg/kg, po measured at 12 hrs	Mus musculus	1		organism-based format	Scientific Literature
18.	ALA4031835	A	Ratio of unbound drug level in C57BL/6 mouse brain to blood at 25 mg/kg, po	Mus musculus	1		organism-based format	Scientific Literature
19.	ALA4031836	F	In vivo inhibition of LRRK2 phosphorylation in brain of C57BL/6 mouse assessed as phosphorylated LRRK2-ser935 levels at 25 mg/kg, po measured at 24 hrs post dose by Western blot analysis	Mus musculus	1		organism-based format	Scientific Literature
20.	ALA4031837	B	Inhibition of recombinant human TYK2 JH1 catalytic domain (887 to 1187 residues) expressed in mammalian expression system at 1 uM by KINOMEScan assay relative to control	Homo sapiens	1		assay format	Scientific Literature