Basic Information
ID: ALA3705221
Type: Binding
Description: ATP Depletion Assay: The activity of PIM2 is measured using a luciferase-luciferin based ATP detection reagent to quantify ATP depletion resulting from kinase-catalyzed phosphoryl transfer to a peptide substrate. Compounds to be tested are dissolved in 100% DMSO and directly distributed into white 384-well plates at 0.5 ul per well. To start the reaction, 10 ul of 10 nM Pim2 kinase and 20 uM BAD peptide (RSRHSSYPAGT-OH) in assay buffer (50 mM HEPES pH 7.5, 5 mM MgCl2, 1 mM DTT, 0.05% BSA) is added into each well. After 15 minutes, 10 ul of 8 uM ATP in assay buffer is added. Final assay concentrations are 5 nM PIM2, 4 uM ATP, 10 uM BAD peptide and 2.5% DMSO. The reaction is performed until approximately 50% of the ATP is depleted, then stopped with the addition of 20 ul KinaseGlo Plus (Promega Corporation) solution. The stopped reaction is incubated for 10 minutes and the remaining ATP detected via luminescence on the Victor2 (Perkin Elmer).
Format: BAO_0000357
Organism: Homo sapiens
Target: Serine/threonine-protein kinase PIM2(ALA4523)
Document: ALA3638844
BindingDB Source: 6370
Activity Charts
Bioactivity
Activity Types for Assay ALA3705221
IC50
Compound Summary
| Parent Molecular Weight | ALogP | Polar Surface Area |
|---|---|---|
| 429.47 | 5.61 | 5.61 |
| 448.48 | 3.61 | 3.61 |
| 448.48 | 3.61 | 3.61 |
| 447.49 | 4.22 | 4.22 |
| 461.52 | 4.46 | 4.46 |
| 476.53 | 4.66 | 4.66 |
| 432.48 | 4.64 | 4.64 |
| 447.49 | 3.58 | 3.58 |
| 450.47 | 4.59 | 4.59 |
| 450.47 | 4.59 | 4.59 |
| 446.51 | 5.03 | 5.03 |
| 433.47 | 4.04 | 4.04 |
| 464.50 | 4.84 | 4.84 |
| 446.51 | 4.89 | 4.89 |
| 446.51 | 4.89 | 4.89 |
| 403.52 | 3.82 | 3.82 |
| 432.48 | 4.64 | 4.64 |
| 414.49 | 4.50 | 4.50 |
| 479.61 | 5.49 | 5.49 |