Reduction of graphene oxide is one of the most promising strategies for obtaining bulk quantities of graphene-like materials. In this study, graphene oxide was deposited on SiO2 and reduced by annealing at 500 K under vacuum conditions (5 × 10−1 Pa). Here, graphene oxide films as well as their chemical changes upon heating were characterized in depth by X-ray photoelectron spectroscopy, Raman spectroscopy, and scanning electron and atomic force microscopies. From the chemical point of view, the as prepared graphene oxide films presented a large quantity of oxidized functional groups that were reduced to a large extent upon heating. Moreover, residual oxidized sulfur species that originated during the synthesis of graphene oxide (GO) were almost completely removed by heating while nitrogen traces were integrated into the carbon framework. On the other hand, regarding structural considerations, reduced graphene oxide films showed more homogeneity and lower roughness than graphene oxide films.