Nonfouling surfaces are essential for many biomedical applications, such as diagnostic biosensors and blood- or tissue-contacting implants. In this study, we demonstrate a simple one-step method to introduce dextran onto various substrates based on dopamine polymerization. It has been shown for the first time that dextran molecules could be incorporated into a dopamine polymerization product via mixing dextran with dopamine in a slightly alkaline solution. The codeposited film was characterized by X-ray photoelectron spectroscopy (XPS), the water contact angle, ellipsometry, and atomic force microscopy (AFM). Results reveal that it is possible to control the thickness and surface roughness via the deposition time and deposition repeat cycles. Furthermore, quartz crystal microbalance (QCM) measurements show that the dextran-modified surface inhibits protein adhesion. In addition, cell attachment has been significantly inhibited on dextran-modified surfaces even after exposure to water for as long as 2 months. The described dopamine-assisted dextran modification represents a simple and universal method for nonfouling surface preparation and can be potentially applied to improve the performance of various medical devices and materials.