Basic Information
ID: ALA3888634
Type: Binding
Description: Time Resolved Fluorescence Energy Transfer (TR-FRET) Assay: Assay 1: The test is performed in white 384-well plates (Greiner Bio-One, reference 781207) in a total volume of 60 μL by adding 1 μL of compounds tested at different concentrations diluted in 100% DMSO (1.7% final DMSO concentration) in reaction buffer (PBS, 125 mM NaCl, 0.001% Novexin (consists of carbohydrate polymers), designed to increase the solubility and stability of proteins; Expedeon Ltd., Cambridgeshire, United Kingdom), 0.01% Gelatin, 0.01% 0.2%, Pluronic F-127 (block copolymer from ethylenoxide and propyleneoxide), 1 mM DTT). After addition of 1.25 nM MDM2-biotinylated or 2.5 nM MDM4-biotinylated (internal preparations), and 0.625 nM Europium labeled streptavidin (Perkin Elmer), the solution is pre-incubated for 15 minutes at room temperature, then 10 nM Cy5-p53 peptide (internal preparation) is added before an incubation at room temperature for 15 minutes prior to reading the plate. For measurement of samples, a Victor II microplate reader (Perkin Elmer) is used with the following settings: Excitation 340 nm, Emission Donor 620 nm and Emission Acceptor 665 nm.
Activity Charts
Bioactivity
Activity Types for Assay ALA3888634
IC50
Compound Summary
| Parent Molecular Weight | ALogP | Polar Surface Area |
|---|---|---|
| 523.03 | 5.94 | 5.94 |
| 496.01 | 4.62 | 4.62 |
| 532.00 | 4.71 | 4.71 |
| 554.43 | 5.30 | 5.30 |
| 608.52 | 6.76 | 6.76 |
| 580.49 | 7.61 | 7.61 |
| 500.99 | 4.60 | 4.60 |
| 553.06 | 4.55 | 4.55 |
| 550.10 | 5.91 | 5.91 |
| 541.43 | 8.39 | 8.39 |
| 481.98 | 4.49 | 4.49 |
| 580.49 | 7.61 | 7.61 |
| 511.02 | 5.23 | 5.23 |
| 501.97 | 4.70 | 4.70 |
| 544.02 | 4.56 | 4.56 |
| 562.50 | 7.47 | 7.47 |
| 493.05 | 6.56 | 6.56 |
| 576.52 | 7.78 | 7.78 |
| 525.05 | 5.35 | 5.35 |
| 523.44 | 8.25 | 8.25 |
| 509.01 | 3.94 | 3.94 |
| 505.45 | 8.11 | 8.11 |