Graphene, as a steady two dimensional (2D) carbon material, possesses intriguing physical and chemical properties, which arouses great interests of scientists for its applications in enormous fields. In particular, graphene and graphene oxide have been widely used for drug delivery and DNA detection based on π-π stacking and hydrophobic interactions. Besides, graphene with fluorescent molecules or nanoparticles and graphene quantum dots have also been frequently applied as fluorescent probe. In this article, advances of graphene and graphene oxide on biomedical applications will be highlighted from the perspective of biomolecular interaction, cell imaging, drug delivery, and toxicity.