Soybeans (Glycine max L.) are one of the most important sources of food allergens, along with cow's milk and eggs; they are recognized as the "three major allergens" of children, which seriously affect children's health. ss-conglycinin is the major soybean allergen, and the alpha subunit of beta-conglycinin, Gly m Bd 60K, is an important allergen in susceptible populations; it was the earliest recognized major allergenic protein. Before this work, we confirmed that the processing treatment reduced the antigenicity of beta-conglycinin, and we located the destroyed antigenic sites of Gly m Bd 60K after three processing technologies. In this paper, we used overlapping peptide technology to further study the destroyed antigenic sites of Gly m Bd 60 K. Finally, one epitope that contained both conformational and a linear IgE epitope was found with an amino acid sequence of (380)EGQQQGEQRLQ(390). Alanine scanning of this fragment documented that Q382, Q383, G385, and Q387 were the critical amino acids of this epitope.