This paper studies the effects of different extraction solution, ratio of sampleweight to extractionsolution and concentrations of reduction reagent dithiothreito (DTT) on extraction efficiency and quality of electrophoresis of prolamin from seeds of Sorghum sudanense. Contribution of the gel concentration and loading dosage to the results of SDSPAGE  were analyzed by evaluating the electrophoresis. The results show that 70% propylalcohol supplemented with 0.2% DTT is the best extraction solution and the ratio of sampleweight to extractionsolution 1∶8 (g/mL) can give optimal results. SDSPAGE of 12% produces the best clearance for electrophoresis under a loading appropriate sample. Established condition based on prolamin from seeds of Sorghum sudanense can be applied to isolation and electrophoresis for prolamins from several other gramineae seeds with good results.