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Assay Detail

CHEMBL3887297

Review assay metadata, readout intent, target linkage, and publication context from the same page.

Binding
AKT Enzyme Assay: A TTP Mosquito liquid handling instrument is used to place 125 nl of the appropriate concentration of inhibitor in 100% DMSO (for a dose response curve calculation) into each well of a 384-well plate. To this reaction components are added to a final volume of 12.5 .mu.l: 0.1 ng/.mu.l His-AKT (Full Length), (Invitrogen, Part # P2999, Lot #641228C). 160 uM ATP (Fluka, 02055) 1 mM DTT (Sigma, D0632) 1 mM MgCl2 (Sigma, M1028) 1 .mu.M substrate peptide (sequence FITC-AHA-GRPRTSSFAEG-NH2), synthesized by Tufts Peptide Synthesis service. 100 mM HEPES pH 7.5 (Calbiochem, 391338)0.015% Brij-35 (Sigma, B4184)The reaction is incubated for 90 min at 25 C, and then stopped by the addition of 70 .mu.l of Stop buffer (100 mM HEPES pH 7.5, 0.015% Brij-35, 10 mM EDTA (Sigma, E7889)).The plate is read on a Caliper LC 3000 in an Off-Chip mobility shift assay format, using the following parameters for a 12-sipper chip: screening pressure -2.3 psi, upstream voltage -500, and downstream voltage -3000.
372
Total Activities
371
Compounds Tested
1
Activity Types
0
Assay Parameters

Assay Information

Assay Type Binding
Organism Homo sapiens
Confidence 8 — Homologous single protein target
Curated By Autocuration

Target

RAC-alpha serine/threonine-protein kinase (CHEMBL4282)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Imidazole amines as modulators of kinase activity
(2015)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 372 7.50 10.00

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3899716 1 10.00
CHEMBL3966806 1 9.82
CHEMBL3936990 1 9.52
CHEMBL3906990 1 9.40
CHEMBL3943127 1 9.28
CHEMBL3980521 1 9.15
CHEMBL3927358 1 9.15
CHEMBL3986943 1 9.10
CHEMBL3960355 1 9.10
CHEMBL3971598 1 9.05
CHEMBL3919089 1 9.00
CHEMBL3941283 1 9.00
CHEMBL3966203 1 8.89
CHEMBL3954981 1 8.85
CHEMBL3890352 1 8.85
CHEMBL3948454 1 8.85
CHEMBL3899490 1 8.82
CHEMBL3967010 1 8.82
CHEMBL3894849 1 8.82
CHEMBL3912471 1 8.82
CHEMBL3941451 1 8.80
CHEMBL3983282 1 8.80
CHEMBL3949795 1 8.77
CHEMBL3944201 1 8.77
CHEMBL3948085 1 8.77
CHEMBL3969016 1 8.77
CHEMBL3980219 1 8.74
CHEMBL3940867 1 8.72
CHEMBL3965271 1 8.72
CHEMBL3984164 1 8.72

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3899716 IC50 = 0.1 nM 10.00
CHEMBL3966806 IC50 = 0.15 nM 9.82
CHEMBL3936990 IC50 = 0.3 nM 9.52
CHEMBL3906990 IC50 = 0.4 nM 9.40
CHEMBL3943127 IC50 = 0.52 nM 9.28
CHEMBL3927358 IC50 = 0.7 nM 9.15
CHEMBL3980521 IC50 = 0.7 nM 9.15
CHEMBL3960355 IC50 = 0.8 nM 9.10
CHEMBL3986943 IC50 = 0.8 nM 9.10
CHEMBL3971598 IC50 = 0.9 nM 9.05
CHEMBL3919089 IC50 = 1.0 nM 9.00
CHEMBL3941283 IC50 = 1.0 nM 9.00
CHEMBL3966203 IC50 = 1.3 nM 8.89
CHEMBL3948454 IC50 = 1.4 nM 8.85
CHEMBL3954981 IC50 = 1.4 nM 8.85
CHEMBL3890352 IC50 = 1.4 nM 8.85
CHEMBL3912471 IC50 = 1.5 nM 8.82
CHEMBL3899490 IC50 = 1.5 nM 8.82
CHEMBL3894849 IC50 = 1.5 nM 8.82
CHEMBL3967010 IC50 = 1.5 nM 8.82
CHEMBL3941451 IC50 = 1.6 nM 8.80
CHEMBL3983282 IC50 = 1.6 nM 8.80
CHEMBL3948085 IC50 = 1.7 nM 8.77
CHEMBL3944201 IC50 = 1.7 nM 8.77
CHEMBL3969016 IC50 = 1.7 nM 8.77
CHEMBL3949795 IC50 = 1.7 nM 8.77
CHEMBL3980219 IC50 = 1.8 nM 8.74
CHEMBL3895898 IC50 = 1.9 nM 8.72
CHEMBL3965271 IC50 = 1.9 nM 8.72
CHEMBL3984164 IC50 = 1.9 nM 8.72
CHEMBL3940867 IC50 = 1.9 nM 8.72
CHEMBL3961056 IC50 = 2.0 nM 8.70
CHEMBL3904291 IC50 = 2.0 nM 8.70
CHEMBL3910789 IC50 = 2.1 nM 8.68
CHEMBL3916178 IC50 = 2.1 nM 8.68
CHEMBL3928772 IC50 = 2.1 nM 8.68
CHEMBL3891980 IC50 = 2.2 nM 8.66
CHEMBL3986793 IC50 = 2.2 nM 8.66
CHEMBL3977147 IC50 = 2.2 nM 8.66
CHEMBL3981085 IC50 = 2.3 nM 8.64
CHEMBL3903522 IC50 = 2.3 nM 8.64
CHEMBL3967750 IC50 = 2.3 nM 8.64
CHEMBL3972479 IC50 = 2.3 nM 8.64
CHEMBL3927877 IC50 = 2.3 nM 8.64
CHEMBL3894578 IC50 = 2.3 nM 8.64
CHEMBL3912769 IC50 = 2.4 nM 8.62
CHEMBL3938454 IC50 = 2.4 nM 8.62
CHEMBL3933597 IC50 = 2.4 nM 8.62
CHEMBL3896910 IC50 = 2.5 nM 8.60
CHEMBL3919834 IC50 = 2.5 nM 8.60