The genus Hyoscyamus belongs to the tribe Hyoscyameae Miers of Solanaceae family. Variation in protein bands elaborates the relationship among the collections from various geographical regions. In this study the seed storage protein diversity of 19 accessions of Hyoscyamus (H. niger, H. reticulatus and H. pusillus) from West Azerbaijan (Iran) was investigated according to the Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) method. Seed protein was analyzed through slab type SDS-PAGE using 15% polyacrylamide gels. The similarity matrix was computed by using Jaccard΄s similarity coefficients, based on polymorphic bands, and dendrogram established through UPGMA cluster analysis. Genetic similarity ranged from 0.07 to 0.87, which indicates high genetic distance at interspecies level. The plotted dendrogram revealed two major clusters. The result of this study indicates that the accessions of Hyoscyamus could effectively be differentiated on the basis of polymorphism, detected by protein bands pattern. Therefore, seed protein profiles seem to be rapid and reliable method to detect inter and intra-specific variation in Hyoscyamus landraces in relation to geographical origin for future breeding programs.