Herein, we describe a study aimed at synthesizing mixed-monolayer-protected CdS nanoparticles and investigating the reactivity of surface-bound functional groups in order to facilitate the immobilization of nanoparticles on a solid substrate as well as the construction of a three-dimensional nanocomposite. CdS nanoparticles initially prepared by the reverse micelle method were used to modify nanoparticle surfaces with 1-decanethiol molecules by ligand exchange. Subsequently, 11-mercapto-1-undecanol was partially incorporated by a place exchange reaction, thereby providing stable, mixed-monolayer-protected CdS nanoparticles. The nanoparticles obtained at each step were characterized by FT-IR and UV-vis spectroscopy, transmission electron microscopy, and elemental analysis. The reactivity of surface hydroxyl groups was verified by a reaction with isocyanate-bearing molecules that provide carbamate bonds in high yields at ambient temperature. The obtained mixed-monolayer-protected nanoparticles were also successfully immobilized on a glass substrate through a carbamate-bond-forming reaction that could be further utilized for multilayer construction in a layer-by-layer fashion.