| Assay Method Information | |
| | Neurotensin Scintillation Proximity Assay |
| Description: | Compound affinity was determined by measuring the displacement of[3H]-neurotensin or [125I]-neurotensin binding to hSortilin in SPA format. Total volume of 40 μl in 50 mM HEPES pH 7.4 assay buffer containing 100 mM NaCl, 2.0 mM CaCI2, 0.1% BSA and 0.1% Tween-20. Compound pre-incubation for 30 minutes at room temperature with 150 nM of 6his-Sortilin before 5 nM [3H]-Neurotensin or [125I]-neurotensin and Ni chelate imaging beads (Perkin Elmer) were added, after 6 hours plate was read on a ViewLux with 360 s exposure time. Dose-response evaluation of compounds was performed with 8 concentrations of drugs (covering 3 decades). IC50 values were calculated by nonlinear regression using the sigmoid concentration-response (variable slope) using CDD Vault software. |
| Affinity data for this assay | |
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