The invention belongs to the field of immobilization of proteins, and relates to preparation and application of a dopamine functional magnetic nano-carrier. The preparation comprises the following steps: firstly, dissolving FeSO4.4H2O and FeCl3.6H2O into distilled water, high-speed stirring and under protection of nitrogen, adding ammonia water of 25% NH4OH, and adding oleic acid; secondly, repeatedly cleaning black precipitates obtained by a stirring reaction and then using a strong magnet to perform precipitation separation, and drying to obtain oleic acid magnetic nano-particles; and finally, adding the oleic acid magnetic nano-particles into a dopamine hydrochloric acid solution, after the reaction is over, using a magnet to separate, cleaning and drying obtained deep brown precipitates, and then the dopamine functional magnetic nano-carrier can be obtained. According to the preparation and application of the dopamine functional magnetic nano-carrier provided by the invention, dopamine hydrochloride is adopted for modifying magnetic oleic acid nano-particles, so that the surface of a magnetic nano-particle has a plentiful of quinonyls and amidogen functional groups, thus proteins can be effectively immobilized; besides, the surface of the carrier has a layer of soft poly dopamine oxide films, the surface area is larger, and the preparation method is simple.