The relic “the sack of Saint Francesco” has for the first time been investigated by scientific means. The sack is kept at the Franciscan Friary of Folloni near Montella in southern Italy. According to legend, the sack appeared on the doorstep of the Friary in the winter of 1224 containing bread sent from St Francesco (St Francis of Assisi), who at that time was in France. The bread was allegedly brought to the friary by an angel. We analyzed samples of the sack to obtain a radiocarbon (14C) date and to search for any remaining traces of bread. The 14C date yielded a calibrated age range of AD 1220–1295 (2σ), which places the textile in the right timeframe according to the legend. Chemical analysis by gas-chromatography with mass spectrometric detection (GC-MS) revealed the presence of ergosterol (5, 7, 22-ergostatrien-3b-ol), a known biomarker of brewing, baking, or agriculture. In this paper we have further substantiated the validity of ergosterol as a biomarker for the past presence of bread. It appears that there is a fine correspondence between the Franciscan legend and the two most decisive scientific methods relevant for analyzing the sack. Although it is not proof, our analysis shows that the sack indeed could be authentic.