At the University of Berlin, Graefe studied philosophy, logic, natural sciences and anatomy, under notable names such as Dove, H. Rose, Müller, and Schlemm, eventually obtaining his medical doctorate in 1847. He continued his studies at Prague, Paris, Vienna and London, and having devoted special attention to ophthalmology, in 1850, he began to practice as an oculist in Berlin. Here, he founded a private institution for the treatment of eyes. During the same year, he received his habilitation with the thesis "Über die Wirkung der Augenmuskeln".