| Assay Method Information | |
| | In Vitro Binding Screening Assay |
| Description: | Equipment: MicroBeta2 LumiJET system, PerkinElmer. Method: Chem-1 cell membranes stably expressing human GnRHR were resuspended in HEPES at pH 7.4. 6 μg of human GnRHR cell membranes were added to each well and incubated with 0.05 nM [125I]-[D-Trp6]-LH-RH and each of the test compounds at 25° C. for 60 min. The compounds were each dissolved in DMSO, with test concentrations being 500 nM, 62.5 nM, 7.81 nM, 0.97 nM, 0.12 nM, 15 pM, and 1.9 pM. 1 pM [D-Trp6]-LH-RH was used in the non-specific binding assay. Cell membranes were collected by filtration under vacuum and washed before liquid scintillation. The binding IC50 and IC90 of the compounds were calculated using Prism. |
| Affinity data for this assay | |
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