Basic Information
ID: ALA3887864
Type: Binding
Description: Luminex Assay: The cellular assay for the determination of the PDK1 kinase activity is carried out as a Luminex assay in the 96-well format. PC3 cells are sown at 20,000 cells per well in 100 ul of medium (45% of RPMI1460/45% of Ham's F12/10% of FCS) and incubated on the following day for 30 min with a serial dilution of the test substance (7 concentrations) under serum-free conditions. The cells are subsequently lysed using 90 ul of lysis buffer (20 mM tris/HCl pH 8.0, 150 mM NaCl, 1% of NP40, 10% of glycerol, 1% of phosphatase inhibitor I, 1% of phosphatase inhibitor II, 0.1% of protease inhibitor cocktail III, 0.01% of benzonase) per well, and the lysates are separated off from insoluble cell constituents by means of centrifugation through a 96-well filter plate (0.65 um). The lysates are incubated overnight at 4° C. with shaking with Luminex beads to which an anti-total PKB antibody is coupled. The detection is carried out on the following day by addition of a phospho-T308-PKB antibody and a species-specific peroxidase-labelled secondary antibody.
Organism: Homo sapiens
Cell type: PC-3
Activity Charts
Bioactivity
Activity Types for Assay ALA3887864
IC50
Compound Summary
| Parent Molecular Weight | ALogP | Polar Surface Area |
|---|---|---|
| 414.45 | 3.91 | 3.91 |
| 409.50 | 4.33 | 4.33 |
| 407.48 | 4.08 | 4.08 |
| 426.48 | 3.13 | 3.13 |
| 486.54 | 2.50 | 2.50 |
| 435.54 | 4.86 | 4.86 |
| 469.98 | 5.52 | 5.52 |
| 430.90 | 4.42 | 4.42 |
| 414.45 | 3.91 | 3.91 |
| 430.90 | 4.42 | 4.42 |
| 396.46 | 3.77 | 3.77 |
| 410.49 | 4.16 | 4.16 |
| 444.93 | 4.81 | 4.81 |
| 424.51 | 4.03 | 4.03 |
| 488.98 | 4.83 | 4.83 |
| 460.93 | 3.79 | 3.79 |
| 426.46 | 3.84 | 3.84 |
| 454.51 | 4.88 | 4.88 |
| 458.96 | 5.47 | 5.47 |
| 418.49 | 3.01 | 3.01 |
| 470.54 | 3.15 | 3.15 |
| 481.52 | 2.37 | 2.37 |
| 488.94 | 4.27 | 4.27 |
| 462.46 | 3.41 | 3.41 |
| 444.47 | 3.27 | 3.27 |
| 424.47 | 3.53 | 3.53 |
| 513.58 | 3.61 | 3.61 |
| 474.96 | 4.44 | 4.44 |
| 435.50 | 4.05 | 4.05 |
| 399.43 | 4.33 | 4.33 |
| 439.53 | 3.49 | 3.49 |
| 453.51 | 3.02 | 3.02 |
| 425.47 | 4.22 | 4.22 |
| 481.56 | 3.67 | 3.67 |
| 439.52 | 4.30 | 4.30 |
| 486.97 | 4.77 | 4.77 |