Basic Information
ID: ALA3706142
Type: Binding
Description: Radioligand Competition Binding Assay: The affinity of the test compounds was determined by radioligand competition binding assay, using the known compound [3H]Ro-15-1788 (Flumazenil) (Perkin Elmer, 85.4 Ci/mmol) and the human recombinant GABA A receptor containing the alpha2, beta2, and gamma3 subunits.Membranes were prepared from HEK cells expressing hGABA A alpha2beta2- gamma3 receptor, and validated to ascertain protein concentration, receptor expression and to determine the Kd of the flumazenil as well as the Ki of a standard set of compounds before being used to test new compounds.The assay was carried out in 96 well plates; testing compounds using a 10 point semi-log dilution range from 19 uM top concentration. 100 ul of radioligand and 100 ul of membrane in 50 mM Tris-HCI and 0.05% F127 with 1 ul of test compound was incubated for 2 hours to allow the reaction to achieve equilibrium, and then harvested onto filter plates, dried and counted on a TopCount NXT. The data was analysed, and the Ki values were presented.
Cell type: HEK293
Activity Charts
Bioactivity
Activity Types for Assay ALA3706142
Ki
Compound Summary
| Parent Molecular Weight | ALogP | Polar Surface Area |
|---|---|---|
| 424.50 | 4.50 | 4.50 |
| 414.44 | 3.86 | 3.86 |
| 397.44 | 2.97 | 2.97 |
| 440.50 | 4.12 | 4.12 |
| 449.51 | 4.37 | 4.37 |
| 466.54 | 4.65 | 4.65 |
| 452.51 | 4.26 | 4.26 |
| 461.52 | 4.69 | 4.69 |
| 422.49 | 4.43 | 4.43 |
| 454.50 | 4.96 | 4.96 |
| 436.51 | 4.60 | 4.60 |
| 436.51 | 4.82 | 4.82 |
| 452.51 | 4.44 | 4.44 |
| 424.50 | 4.50 | 4.50 |
| 424.50 | 4.67 | 4.67 |
| 411.46 | 3.53 | 3.53 |
| 425.49 | 3.79 | 3.79 |
| 442.49 | 4.81 | 4.81 |
| 463.54 | 4.93 | 4.93 |
| 467.53 | 4.07 | 4.07 |
| 480.57 | 5.22 | 5.22 |
| 440.50 | 3.61 | 3.61 |
| 456.96 | 4.64 | 4.64 |
| 452.54 | 3.47 | 3.47 |
| 447.52 | 4.53 | 4.53 |
| 438.51 | 3.69 | 3.69 |
| 410.47 | 4.11 | 4.11 |
| 422.49 | 4.04 | 4.04 |
| 411.46 | 3.23 | 3.23 |
| 396.45 | 3.72 | 3.72 |
| 442.49 | 4.64 | 4.64 |
| 435.48 | 3.98 | 3.98 |
| 426.47 | 3.73 | 3.73 |
| 454.53 | 4.51 | 4.51 |
| 460.48 | 5.05 | 5.05 |
| 428.46 | 4.25 | 4.25 |
| 378.46 | 3.58 | 3.58 |
| 390.47 | 3.51 | 3.51 |
| 423.47 | 3.54 | 3.54 |
| 408.46 | 4.04 | 4.04 |
| 440.48 | 4.57 | 4.57 |
| 466.54 | 4.83 | 4.83 |
| 438.48 | 4.05 | 4.05 |
| 447.50 | 4.30 | 4.30 |
| 456.52 | 5.20 | 5.20 |
| 468.55 | 5.07 | 5.07 |
| 440.50 | 4.29 | 4.29 |
| 458.95 | 5.33 | 5.33 |
| 454.53 | 4.68 | 4.68 |
| 449.51 | 4.55 | 4.55 |