Using graphene oxide as substrate and stabilizer for the silver nanoparticles, silver nanoparticles-graphene oxide (Ag NPs/GO) composites with different Ag loading were synthesized through a facile solution-phase method. During the synthesis process, AgNO3 on GO matrix was directly reduced by NaBH4. The structure characterization was studied through X-ray diffraction (XRD), atomic force microscopy (AFM), high-resolution transmission electron microscope (HRTEM), ultraviolet-visible spectroscopy (UV-Vis), and selected area electron diffraction (SAED). The results show that Ag nanoparticles (Ag NPs) with the sizes ranging from 5 to 20 nm are highly dispersed on the surfaces of GO sheets. The shape and size of the Ag NPs are decided by the volume of initial AgNO3 solution added in the GO. The antibacterial activities of Ag NPs/GO nanocomposites were investigated and the result shows that all the produced composites exhibit good antibacterial activities against Gram-negative (G−) bacterial strain Escherichia coli (E. coli) and Gram-positive (G+) strain Staphylococcus aureus (S. aureus). Moreover, the antibacterial activities of Ag NPs/GO nanocomposites gradually increased with the increasing of volume of initial AgNO3 solution added in the GO and this improvement of the antibacterial activities results from the combined action of size effect and concentration effect of Ag NPs in Ag NPs/GO nanocomposites.