Graphene oxide (GO) is a type of two-dimensional nanomaterial with a single-atom thickness. GO sheets contain pristine regions, oxidized regions, and a small fraction of holes. By stacking GO sheets together, a GO membrane can be fabricated with sufficient mechanical strength. The interlayer nanocapillary network formed from connected interlayer spaces, together with the gaps between the edges of noninterlocked neighboring GO sheets and cracks or holes of the GO sheet, provides passage for molecules or ions to permeate through the GO membrane in an aqueous solution. The characteristics of molecules or ions (e.g., their size, charge, and the interaction with the GO membrane) affect the separation performance of the GO membrane. The contribution of gaps between neighboring GO sheets for separation can be adjusted by changing the GO sheet size and the GO membrane thickness. The interlayer space of the GO membrane can be adjusted by changing the water pH and modifying or reducing the GO sheets to obtain the desired separation performance. The production of the GO membrane is easily scalable and relatively inexpensive, indicating that the GO membrane has promising potential for applications such as water treatment, desalination, anticorrosion, chemical resistance, and controlled release coatings.