Pyrene and its derivatives became a standard tool for the immobilization of bioentities on nanostructured sp 2 carbon such as graphene and carbon nanotubes (CNTs) for bioelectrochemical applications. Stable π-stacking interaction between these compounds can be formed under mild conditions without affecting sensitive anchor functions of pyrene or introducing defect groups on the carbon material. Pyrene has further beneficial properties for bioelectrochemical studies. Pyrene itself can form supramolecular inclusion complexes with β-cyclodextrin and can even interact with the hydrophobic domain of a laccase enzyme allowing oriented immobilization. Pyrene can further act as cross-linker, be electropolymerized to reinforce the formed layer, and also be electro-oxidized becoming an efficient redox mediator for the electron transfer with a FAD co-factor. This versatility of pyrene and its derivatives for bioelectrochemical applications are focused here beside some original anchoring methods for biomolecules.