| Assay Method Information | |
| | Microscale Thermophoresis Assay |
| Description: | The binding affinity of COMPOUND I and compounds of the invention to human sRAGE (the soluble extracellular domain of the human receptor for advanced glycation endproducts) was measured using microscale thermophoresis.Microscale Thermophoresis System (Monolith NT.115 Pico, NanoTemper, Inc) was used to determine the binding affinity of COMPOUND I and metabolites M1, M2, M3, M5, M6 and M7 to recombinant human sRAGE. sRAGE was labeled with NT-647-NHS fluorescence dye. Around 5 nM sRAGE was subsequently added to compounds at concentrations from 600 nM to 20 pM and incubated for 5 minutes at 25° C. in buffer containing 25 mM Hepes (pH 7.5), 5 mM CaCl2), 5 mM MgCl2, 50 mM NaCl, 0.05% Tween20 and 1% DMSO before proceeding to microscale thermophoresis. |
| Affinity data for this assay | |
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