Anandamide (arachidonoylethanolamide, AEA) acts as endogenous agonist of both cannabinoidand vanilloid receptors. During the last two decades, its metabolic pathways and biologicalactivity have been extensively investigated and relatively well-characterized. In contrast, theeffective nature and mechanism of AEA transport remain at present a controversial and stillunsolved issue. Here we report the characterization of a biotinylated analogue of AEA (b-AEA),that has the same lipophilicity of the parent compound. In addition, by means of biochemicalassays and fluorescence microscopy, we show that b-AEA is accumulated inside the cells in away superimposable on that of AEA. Conversely, b-AEA does not interact nor interfere with theother components of the endocannabinoid system, i.e. type-1 and type-2 cannabinoid receptors,vanilloid receptor, AEA synthetase (NAPE-PLD) or AEA hydrolase (FAAH). Taken together,our data suggest that b-AEA could be a very useful probe for visualizing the accumulation andintracellular distribution of this endocannabinoid.[...]