Electrospinning is a very versatile technique, which holds great potentials for several clinical applications. The ability to produce biocompatible nanofibres mimicking the extracellular matrix of the body in combination with both the large surface area and the adsorption/release properties of nanofibres makes drug loaded electrospun fibres very promising for both drug delivery and tissue engineering purposes. An interesting type of molecules to incorporate into electrospun nanofibres are antimicrobial peptides (AMPs) due to their characteristic mode of action both as antimicrobial- and as immunological agents. The focus of the presented work was on the release properties and the loading density of the synthetic AMP fluorescein labelled inverse-Crabrolin (iCR-fluor) incorporated into electrospun nanofibres of poly(epsilon-caprolactone) (PCL). The release properties were compared to the release properties of fluorescein and tetracycline hydrochloride. Furthermore, the antimicrobial effect of the different loading agents was evaluated both before and after release from the fibres, where only tetracycline hydrochloride was found to retain its activity. The loading density of fluorescein and iCR-fluor was investigated with deconvolution fluorescence microscopy. iCR-fluor followed a linear release profile with a significantly slower release kinetics than tetracycline hydrochloride and fluorescein. After the first 60 min, approximately 85% of both fluorescein and tetracycline were released, whereas only 40% of iCR-fluor was released. Furthermore, iCR-fluor did not show uniform distribution within the fibres and had an overall lower loading density than fluorescein.