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Assay Detail

CHEMBL3888429

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

Binding
HTRF Biochemical Assay: The reactions employed a common peptide substrate, LCB-EQEDEPEGDYFEWLW-NH2 (in-house). The basic assay protocol is as follows: First, 250 nL of diluted compounds in DMSO were dispensed into the wells of a dry 384-well Black plate (Greiner #781076) using a Labcyte Echo 555 acoustic dispenser. Subsequent reagent additions employed an Agilent Bravo. Next, 18 uL of 1.11x enzyme and 1.11x substrate in 1x assay buffer (Invitrogen kinase buffer # PV3189, 2 mM DTT, 0.05% BSA) were added to the wells and shaken and then preincubated for 30 minutes at room temperature to allow compound binding to equilibrate. After equilibration, 2 uL of 10xATP in 1x assay buffer was added to initiate the kinase reaction and the plates were shaken and then incubated at room temperature for 120 minutes. At the end of the incubation, 20 uL of 2x stop buffer (streptavidin-Dylight 650 (Thermo #84547B/100 mL), Eu-tagged pY20 antibody (Perkin Elmer #AD0067), EDTA, HEPES, and Triton) was added to quench the reaction.
667
Total Activities
620
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

Tyrosine-protein kinase JAK2 (CHEMBL2971)
Type SINGLE PROTEIN
Organism Homo sapiens

Publication

Cyanomethylpyrazole carboxamides as janus kinase inhibitors
(2016)

Activity Statistics

Type Count Avg pChEMBL Best pChEMBL
IC50 667 7.26 10.00

Compounds Tested

Compound Name Phase Activities Best pChEMBL
CHEMBL3965448 1 10.00
CHEMBL3972665 1 10.00
CHEMBL4106834 1 9.70
CHEMBL3947540 2 9.70
CHEMBL3959005 1 9.70
CHEMBL3929513 1 9.30
CHEMBL3922069 1 9.30
CHEMBL3935178 1 9.22
CHEMBL3929641 1 9.22
CHEMBL3896068 1 9.22
CHEMBL3984300 1 9.22
CHEMBL3895272 1 9.22
CHEMBL3942917 1 9.15
CHEMBL3902563 1 9.15
CHEMBL3931250 1 9.15
CHEMBL3931150 1 9.10
CHEMBL3904265 3 9.10
CHEMBL3938031 2 9.10
CHEMBL3935508 1 9.00
CHEMBL3914600 1 9.00
CHEMBL3949472 1 9.00
CHEMBL3933102 1 9.00
CHEMBL3910522 1 9.00
CHEMBL3950253 2 9.00
CHEMBL3892599 2 9.00
CHEMBL3919665 2 9.00
CHEMBL3916939 1 8.70
CHEMBL3932352 1 8.70
CHEMBL3977608 1 8.70
CHEMBL3968473 1 8.70

Activity Data

Compound Name Type Rel. Value Units pChEMBL
CHEMBL3972665 IC50 = 0.1 nM 10.00
CHEMBL3965448 IC50 = 0.1 nM 10.00
CHEMBL4106834 IC50 = 0.2 nM 9.70
CHEMBL3947540 IC50 = 0.2 nM 9.70
CHEMBL3959005 IC50 = 0.2 nM 9.70
CHEMBL3922069 IC50 = 0.5 nM 9.30
CHEMBL3929513 IC50 = 0.5 nM 9.30
CHEMBL3896068 IC50 = 0.6 nM 9.22
CHEMBL3984300 IC50 = 0.6 nM 9.22
CHEMBL3929641 IC50 = 0.6 nM 9.22
CHEMBL3935178 IC50 = 0.6 nM 9.22
CHEMBL3895272 IC50 = 0.6 nM 9.22
CHEMBL3942917 IC50 = 0.7 nM 9.15
CHEMBL3902563 IC50 = 0.7 nM 9.15
CHEMBL3931250 IC50 = 0.7 nM 9.15
CHEMBL3938031 IC50 = 0.8 nM 9.10
CHEMBL3904265 IC50 = 0.8 nM 9.10
CHEMBL3931150 IC50 = 0.8 nM 9.10
CHEMBL3935508 IC50 = 1.0 nM 9.00
CHEMBL3910522 IC50 = 1.0 nM 9.00
CHEMBL3933102 IC50 = 1.0 nM 9.00
CHEMBL3892599 IC50 = 1.0 nM 9.00
CHEMBL3950253 IC50 = 1.0 nM 9.00
CHEMBL3904265 IC50 = 1.0 nM 9.00
CHEMBL3914600 IC50 = 1.0 nM 9.00
CHEMBL3919665 IC50 = 1.0 nM 9.00
CHEMBL3949472 IC50 = 1.0 nM 9.00
CHEMBL3982094 IC50 = 2.0 nM 8.70
CHEMBL3928653 IC50 = 2.0 nM 8.70
CHEMBL3965904 IC50 = 2.0 nM 8.70
CHEMBL3932064 IC50 = 2.0 nM 8.70
CHEMBL3920570 IC50 = 2.0 nM 8.70
CHEMBL3915313 IC50 = 2.0 nM 8.70
CHEMBL3943909 IC50 = 2.0 nM 8.70
CHEMBL3950965 IC50 = 2.0 nM 8.70
CHEMBL3916939 IC50 = 2.0 nM 8.70
CHEMBL3917637 IC50 = 2.0 nM 8.70
CHEMBL3904021 IC50 = 2.0 nM 8.70
CHEMBL3932352 IC50 = 2.0 nM 8.70
CHEMBL3973424 IC50 = 2.0 nM 8.70
CHEMBL3914311 IC50 = 2.0 nM 8.70
CHEMBL3929554 IC50 = 2.0 nM 8.70
CHEMBL3968473 IC50 = 2.0 nM 8.70
CHEMBL3977608 IC50 = 2.0 nM 8.70
CHEMBL3983487 IC50 = 2.0 nM 8.70
CHEMBL3916947 IC50 = 2.0 nM 8.70
CHEMBL3958942 IC50 = 2.0 nM 8.70
CHEMBL3920186 IC50 = 2.0 nM 8.70
CHEMBL3945594 IC50 = 2.0 nM 8.70
CHEMBL3897977 IC50 = 2.0 nM 8.70