Basic Information
ID: ALA3707632
Type: Binding
Description: Kinase Assay: The compounds from the examples below were investigated for their CDK2/cyclin E, CDK1/cyclin B, CDK4/cyclin D1 and CDK7/cyclin H, ERK-2, and PKA inhibitory activity. His6-tagged recombinant human cyclin-dependent kinases CDK1/cyclin B1, CDK2/cyclin E, CDK4 and CDK7/cyclin H were expressed in sf9 cells using a baculovirus expression system. Recombinant cyclin D1 was expressed in E. coli. Proteins were purified by metal chelate affinity chromatography to greater than 90% homogeneity. Kinase assays were performed in 96-well plates using recombinant CDK/cyclins, recombinant active ERK-2 (Upstate Biotechnology), or cyclic AMP-dependent kinase (PKA) catalytic subunit (Calbiochem Cat. 539487). Assays were performed in assay buffer (25 mM beta -glycerophosphate, 20 mM MOPS, 5 mM EGTA, 1 mM DTT, 1 mM Na3VO3, pH 7.4), into which were added 2-4 ug of active enzyme with appropriate substrates (purified histone H1 for CDK2, recombinant GST-retinoblastoma protein (residues 773-928) for CDK4.
Organism: Homo sapiens
Cell type: Sf9
Activity Charts
Bioactivity
Activity Types for Assay ALA3707632
IC50
Compound Summary
| Parent Molecular Weight | ALogP | Polar Surface Area |
|---|---|---|
| 383.50 | 3.38 | 3.38 |
| 397.53 | 3.62 | 3.62 |
| 411.55 | 4.01 | 4.01 |
| 383.50 | 3.38 | 3.38 |
| 383.50 | 3.38 | 3.38 |
| 397.53 | 3.62 | 3.62 |
| 411.55 | 4.01 | 4.01 |
| 383.50 | 3.38 | 3.38 |
| 369.47 | 2.99 | 2.99 |
| 369.47 | 2.99 | 2.99 |