| Assay Method Information | |
| | Surface Plasmon Resonance Assay |
| Description: | SPR was used to confirm and quantify binding of compounds to human STAT6. Biacore instruments from the 8K-series (Cytiva) were used, and measurements were performed at 25° C. Human STAT6 (truncated, aa122-658, with N-terminal His-tag, expressed in insect cells) was immobilized in the active flow cells on a CM5 chip using amine-coupling (default settings) at 10 μg/ml in 10 mM acetate buffer pH 5.5, at a contact time of 120-420 sec. and a flow rate of 10 μl/min. Reference flow cells were deactivated using blank immobilization. PBS-P+ (Cytiva) supplemented with 2% DMSO was used as running buffer, and solvent correction was applied. Compounds were tested by multi-cycle kinetics injections using a contact time of 60 sec., dissociation time of up to 300 sec., and a flow rate of 30 μl/min. Solvent-corrected, reference-subtracted and blank-corrected data was processed and binding isotherms were fitted according to a single report point from each sensorgram (Standard: Late binding (5 sec. before injection end); alternative: Earlier report point according to binding profile). Data fitting was done both with unfixed Rmax and with Rmax fixed to the theoretical value* assuming 1:1 binding to allow processing of data from both weak and strong binders as well as to evaluate binding stoichiometry. |
| Affinity data for this assay | |
|---|---|
| If you find an error in this entry please send us an E-mail | |