Assay Detail
Binding
CHEMBL5730045
Review assay metadata, readout intent, target linkage, and publication context from the same page.
Time-Resolved Fluorescence Resonance energy transfer (TR-FRET) Immunoassay: The assay measures the phosphorylation level of a biotinylated peptide substrate by the ASK1 kinase using HTRF detection. This is a competitive, time-resolved fluorescence resonance energy transfer (TR-FRET) immunoassay, based on HTRF KinEASE-STK manual from Cisbio. Test compound, 1 uM STK3 peptide substrate, 4 nM of ASK1 kinase were incubated with 10 mM MOP buffer, pH. 7.0 containing 10 mM Mg-acetate, 0.025% NP-40, 1 mM DTT, 0.05% BSA and 1.5% glycerol for 30 minutes then 100 uM ATP was added to start the kinase reaction and incubated for 3 hr. Peptide antibody labeled with 1xEu3+ Cryptate buffer containing 10 mM EDTA and 125 nM Streptavidin XL665 were added to stop the reaction and phosphorylated peptide substrate was detected using Envision 2103 Multilabeled reader from PerkinElmer. The fluorescence was measured at 615 nm (Cryptate) and 665 nm (XL665) and a ratio of 665 nm/615 nm was calculated for each well.
135
Total Activities
45
Compounds Tested
3
Activity Types
0
Assay Parameters
Assay Information
| Assay Type | Binding |
| Confidence | 8 — Homologous single protein target |
| Curated By | Autocuration |
Target
RAC-alpha serine/threonine-protein kinase (CHEMBL4282) ↗| Type | SINGLE PROTEIN |
| Organism | Homo sapiens |
Publication
Apoptosis signal-regulating kinase inhibitors
Activity Statistics
| Type | Count | Avg pChEMBL | Best pChEMBL |
|---|---|---|---|
| IC50 | 45 | 8.67 | 9.40 |
| kon | 45 | - | - |
| k_off | 45 | - | - |
Compounds Tested
| Compound | Name | Phase | Activities | Best pChEMBL |
|---|---|---|---|---|
| CHEMBL3969572 | — | — | 3 | 9.40 |
| CHEMBL3975211 | — | — | 3 | 9.05 |
| CHEMBL3922577 | — | — | 3 | 8.96 |
| CHEMBL3902880 | — | — | 3 | 8.92 |
| CHEMBL3893968 | — | — | 3 | 8.85 |
| CHEMBL3976987 | — | — | 3 | 8.82 |
| CHEMBL3930015 | — | — | 3 | 8.77 |
| CHEMBL3917308 | — | — | 3 | 8.77 |
| CHEMBL3962884 | — | — | 3 | 8.77 |
| CHEMBL3905950 | — | — | 3 | 8.74 |
| CHEMBL3979677 | — | — | 3 | 8.74 |
| CHEMBL3925596 | — | — | 3 | 8.74 |
| CHEMBL3897512 | — | — | 3 | 8.74 |
| CHEMBL3909615 | — | — | 3 | 8.72 |
| CHEMBL3948895 | — | — | 3 | 8.72 |
| CHEMBL3935565 | — | — | 3 | 8.72 |
| CHEMBL3924482 | — | — | 3 | 8.72 |
| CHEMBL3954144 | — | — | 3 | 8.72 |
| CHEMBL3946920 | — | — | 3 | 8.70 |
| CHEMBL3953748 | — | — | 3 | 8.70 |
| CHEMBL3961404 | — | — | 3 | 8.68 |
| CHEMBL3961734 | — | — | 3 | 8.68 |
| CHEMBL3908126 | — | — | 3 | 8.66 |
| CHEMBL3953359 | — | — | 3 | 8.66 |
| CHEMBL3898363 | — | — | 3 | 8.66 |
| CHEMBL3893936 | — | — | 3 | 8.66 |
| CHEMBL3986247 | — | — | 3 | 8.64 |
| CHEMBL3926976 | — | — | 3 | 8.64 |
| CHEMBL3904498 | — | — | 3 | 8.64 |
| CHEMBL3908356 | — | — | 3 | 8.64 |
Activity Data
| Compound | Name | Type | Rel. | Value | Units | pChEMBL |
|---|---|---|---|---|---|---|
| CHEMBL3969572 | — | IC50 | = | 0.4 | nM | 9.40 |
| CHEMBL3975211 | — | IC50 | = | 0.9 | nM | 9.05 |
| CHEMBL3922577 | — | IC50 | = | 1.1 | nM | 8.96 |
| CHEMBL3902880 | — | IC50 | = | 1.2 | nM | 8.92 |
| CHEMBL3893968 | — | IC50 | = | 1.4 | nM | 8.85 |
| CHEMBL3976987 | — | IC50 | = | 1.5 | nM | 8.82 |
| CHEMBL3962884 | — | IC50 | = | 1.7 | nM | 8.77 |
| CHEMBL3917308 | — | IC50 | = | 1.7 | nM | 8.77 |
| CHEMBL3930015 | — | IC50 | = | 1.7 | nM | 8.77 |
| CHEMBL3925596 | — | IC50 | = | 1.8 | nM | 8.74 |
| CHEMBL3897512 | — | IC50 | = | 1.8 | nM | 8.74 |
| CHEMBL3905950 | — | IC50 | = | 1.8 | nM | 8.74 |
| CHEMBL3979677 | — | IC50 | = | 1.8 | nM | 8.74 |
| CHEMBL3909615 | — | IC50 | = | 1.9 | nM | 8.72 |
| CHEMBL3948895 | — | IC50 | = | 1.9 | nM | 8.72 |
| CHEMBL3954144 | — | IC50 | = | 1.9 | nM | 8.72 |
| CHEMBL3924482 | — | IC50 | = | 1.9 | nM | 8.72 |
| CHEMBL3935565 | — | IC50 | = | 1.9 | nM | 8.72 |
| CHEMBL3953748 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL3946920 | — | IC50 | = | 2.0 | nM | 8.70 |
| CHEMBL3961734 | — | IC50 | = | 2.1 | nM | 8.68 |
| CHEMBL3961404 | — | IC50 | = | 2.1 | nM | 8.68 |
| CHEMBL3908126 | — | IC50 | = | 2.2 | nM | 8.66 |
| CHEMBL3893936 | — | IC50 | = | 2.2 | nM | 8.66 |
| CHEMBL3898363 | — | IC50 | = | 2.2 | nM | 8.66 |
| CHEMBL3953359 | — | IC50 | = | 2.2 | nM | 8.66 |
| CHEMBL3904498 | — | IC50 | = | 2.3 | nM | 8.64 |
| CHEMBL3986247 | — | IC50 | = | 2.3 | nM | 8.64 |
| CHEMBL3908356 | — | IC50 | = | 2.3 | nM | 8.64 |
| CHEMBL3926976 | — | IC50 | = | 2.3 | nM | 8.64 |
| CHEMBL3899813 | — | IC50 | = | 2.4 | nM | 8.62 |
| CHEMBL3978053 | — | IC50 | = | 2.5 | nM | 8.60 |
| CHEMBL3920406 | — | IC50 | = | 2.5 | nM | 8.60 |
| CHEMBL3903012 | — | IC50 | = | 2.6 | nM | 8.59 |
| CHEMBL3963748 | — | IC50 | = | 2.7 | nM | 8.57 |
| CHEMBL3964706 | — | IC50 | = | 2.7 | nM | 8.57 |
| CHEMBL3954180 | — | IC50 | = | 2.8 | nM | 8.55 |
| CHEMBL3957940 | — | IC50 | = | 2.8 | nM | 8.55 |
| CHEMBL3895494 | — | IC50 | = | 2.8 | nM | 8.55 |
| CHEMBL3953443 | — | IC50 | = | 2.9 | nM | 8.54 |
| CHEMBL3924518 | — | IC50 | = | 3.2 | nM | 8.49 |
| CHEMBL3943452 | — | IC50 | = | 3.2 | nM | 8.49 |
| CHEMBL3893943 | — | IC50 | = | 3.8 | nM | 8.42 |
| CHEMBL3973753 | — | IC50 | = | 4.5 | nM | 8.35 |
| CHEMBL3981380 | — | IC50 | = | 14.2 | nM | 7.85 |
| CHEMBL3904498 | — | k_off | = | - | s-1 | - |
| CHEMBL3908356 | — | k_off | = | - | s-1 | - |
| CHEMBL3909615 | — | kon | = | - | — | - |
| CHEMBL3954144 | — | k_off | = | - | s-1 | - |
| CHEMBL3904498 | — | kon | = | - | — | - |