#	Aladdin ID	Assay Type	Description	Organism	Compounds	Reference	BAO Format	Source
1.	ALA679355	B	Inhibition of ERK2 kinase	Mus musculus	1	ALA1132910	single protein format	Scientific Literature
2.	ALA732076	B	Inhibition of mitogen-activated protein kinase 2	Mus musculus	3	ALA1146654	single protein format	Scientific Literature
3.	ALA732077	B	Inhibition of mouse Mitogen-activated protein kinase 2 at 10 uM	Mus musculus	2	ALA1145082	single protein format	Scientific Literature
4.	ALA827548	B	Inhibition of Mitogen-activated protein kinase/extracellular signal-regulated kinase 2 in P19 cells	Mus musculus	7	ALA1142718	cell-based format	Scientific Literature
5.	ALA898197	B	Increase in ERK2 phosphorylation in mouse B16 cells at 4-8 hrs	Mus musculus	1	ALA1149593	cell-based format	Scientific Literature
6.	ALA939178	B	Inhibition of IGF1-stimulated purified ERK2 phosphorylation in mouse 3T3 cells	Mus musculus	1	ALA1143953	cell-based format	Scientific Literature
7.	ALA939650	B	Reversal of ERK2 phosphorylation in mouse C2C12 at 20 uM in presence of 50 uM LY-294002 after 48 hrs	Mus musculus	1	ALA1156479	single protein format	Scientific Literature
8.	ALA950815	B	Inhibition of mouse MAPK2 at 10 uM	Mus musculus	1	ALA1137681	single protein format	Scientific Literature
9.	ALA1026918	B	Inhibition of GST-fused mouse MAPK2 expressed in Escherichia coli at 10 uM	Mus musculus	1	ALA1158406	assay format	Scientific Literature
10.	ALA1646188	B	Inhibition of mouse MAPK2 at 10 uM after 60 mins by TR-FRET assay	Mus musculus	1	ALA1641539	single protein format	Scientific Literature
11.	ALA1952646	B	Activity at mouse ERK2 receptor at 10 uM	Mus musculus	1	ALA1949601	single protein format	Scientific Literature
12.	ALA2345863	B	Activation of ERK2 in mouse RAW264.7 cells assessed as phosphorylation at Thr185/Tyr187 at 50 uM after 30 mins by Western blotting analysis	Mus musculus	1	ALA2331476	cell-based format	Scientific Literature
13.	ALA2410778	B	Inhibition of mouse ERK2 pre-incubation for 15 mins before substrate addition measured after 60 mins by IMAP-FP assay	Mus musculus	3	ALA2406935	single protein format	Scientific Literature
14.	ALA3591270	B	Binding affinity to ERK2 in mouse fibroblasts assessed as half-life	Mus musculus	1	ALA3588785	single protein format	Scientific Literature
15.	ALA3705054	B	IMAP Assay: Activated ERK2 activity was determined in the IMAP assay format.	Mus musculus	14	ALA3638513	single protein format	BindingDB Database
16.	ALA3706036	B	IMAP-FP Assay: Activated ERK2 activity was determined in an IMAP-FP assay (Molecular Devices). Using this assay format, the potency (IC50) of each compound was determined from a 10 point (1:3 serial dilution, 3 uM starting compound concentration) titration curve using the following outlined procedure. To each well of a black Corning 384-well plate (Corning Catalog #3575), 7.5 mL of compound (3333 fold dilution in final assay volume of 25 uL) was dispensed, followed by the addition of 15 uL of kinase buffer (tween containing kinase buffer, Molecular Devices) containing 0.0133 ng/mL (0.316 nM) of phosphorylated active mERK2 enzyme. Following a 15 minute compound & enzyme incubation, each reaction was initiated by the addition of 10 uL kinase buffer containing 2.45 uM ERK2 IMAP substrate peptides (2.25 uM-unlabeled IPTTPITTTYFFFK-COOH and 200 nM-labeled IPTTPITTTYFFFK-5FAM (5-carboxyfluorescein)-COOH), and 75 uM ATP.	Mus musculus	24	ALA3638680	single protein format	BindingDB Database
17.	ALA3706286	B	Fluorescence Quenching Assay: CADD and biological testing to identify compounds that interact with novel ERK substrate docking sites. The known ERK2 docking sites include the DRS (site 1) and FRS (site 5) regions, whereas predicted docking sites are labeled sites 2-4 (Table 6). One compound, 76, that targets the DRS is being marketed by Calbiochem/EMD Biosciences (Cat#328006) as an ERK inhibitor. Successful inhibitor identification to date has been based on a combination of fluorescence quenching (FQ) of ERK2 wild type and docking site mutants in the presence of test compound.	Mus musculus	8	ALA3638871	single protein format	BindingDB Database
18.	ALA3705760	B	Coupled ERK2 (cERK) Assay: Activity of compounds against inactive ERK2 was tested in a coupled MEK1/ERK2 IMAP assay as follows: Compounds were diluted to 25x final test concentration in 100% DMSO. 14 ul of kinase buffer (10 mM Tris.HCl pH 7.2, 10 mM MgCl2, 0.01% Tween-20, 1 mM DTT) containing 0.4 ng unphosphorylated Mouse ERK2 protein was added to each well of a black 384-well assay plate. 1 ul of 25x compound was added to each well and incubated at room temperature for 30 minutes to allow an opportunity for the compound to bind to the inactive enzyme. DMSO concentration during initial incubation is 6.7%. ERK2 activity was determined to be insensitive to DMSO concentrations up to 20%. ERK2 was then activated and it's kinase activity measured by the addition of 10 ul kinase buffer with the following components (final concentration per reaction): 2 ng active (phosphorylated) human MEK1 protein and 4 uM (total) ERK2 IMAP substrate peptides.	Mus musculus	16	ALA3638826	single protein format	BindingDB Database
19.	ALA3705761	B	IMAP-FP Assay: Condition 2: Activated ERK2 activity was determined in an IMAP-FP assay (Molecular Devices). Using this assay format, the potency (IC50) of each compound was determined from a 10 point (1:3 serial dilution, 3 uM starting compound concentration) titration curve using the following outlined procedure. 7.5 mL of compound (3333 fold dilution in final assay volume of 25 uL) was added to 15 uL of kinase buffer containing 0.0133 ng/mL (0.316 nM) of fully phosphorylated, mouse aERK2 enzyme. Following a 15 minute incubation, each reaction was initiated by the addition of 10 uL kinase buffer containing 2.45 uM ERK2 IMAP substrate peptides (2.25 uM-unlabeled IPTTPITTTYFFFK-COOH and 200 nM-labeled IPTTPITTTYFFFK-5FAM (5-carboxyfluorescein)-COOH), and 75 uM ATP. The final reaction in each well of 25 uL consists of 0.19 nM mouse ERK2, 900 nM unlabeled peptide, 80 nM labeled-peptide, and 30 uM ATP. Phosphorylation reactions were allowed to proceed for 40-45 minutes.	Mus musculus	31	ALA3638826	single protein format	BindingDB Database
20.	ALA3705762	B	IMAP Assay : Condition 1: Activated ERK2 activity was also determined in the IMAP assay format using the procedure outlined above. 1 μl of 25× compound was added to 140 of kinase buffer containing 0.25 ng fully phosphorylated, active Mouse ERK2 protein. Following a 30 minute incubation, the reactions were initiated by addition of 10 μl of kinase buffer containing 1 μM ERK2 IMAP substrate peptide (0.9 μM unlabeled IPTTPITTTYFFFK-CONH2 and 100 nM IPTTPITTTYFFFK(5-carboxyfluorescein)-CONH2) and 30 μM ATP. Reactions proceeded for 30 minutes before termination by addition of 60 μl IMAP detection beads in binding buffer. Plates were read as above after 30 minute binding equilibration. Active compounds were reconfirmed in an independent assay.	Mus musculus	280	ALA3638826	single protein format	BindingDB Database