Brush border membrane vesicles(BBMV)isolated from insect midguts have been widely used to study CrylA binding proteins.Sample preparation is important in two-dimensional electrophoresis(2-DE),so to determine a suitable BBMV preparation method in Helicoverpa armigera for 2-DE,we compared three published BBMV preparation methods mostly used in sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).All mctllods yielded similar types and numbers of binding proteins,but in different quantifies.The Abdul.Rauf and Ellar protocol was the best of the three,but had limitations.Sufficient protein qu antity iS important for research involving limited numbers of insects,such as studies of insect resistance to Bacillus thuringiensis in the field.Consequently,we integrated the three BBMV isolation methods into a single protocol that yielded high quantities of BBMV proteins from H.armigera larval midguts.which proved suitable for 2-DE analysis.