What is the general process used to bind ligands to the beads?
When binding ligands to NHS beads, for the binding reaction, 4 concentrations are prepared (ligand solutions: 0, 0.1, 0.3, 1 mM) in order to determine the optimal concentration. The amount of bound ligands depends on the ligand properties, and varies approximately from several nmol to 100 nmol. An amount of several nmol to several tens of nmol is best for achieving purification results. Too much ligand can cause non-specific adsorption and steric hindrance, which adversely affects affinity purification.
