In order to demonstrate the efficacy of the membrane enrichment method by Urea Wash in the animal model. Kidneys were dissected from 8 weeks old male mice of inbred strain C57BL/6J and flash-frozen in liquid nitrogen followed by storage at -80 °C. Brush border membrane vesicles (BBMV) were prepared. From this study, we demystified the merits of Urea wash methods for membrane enrichment. The membrane enrichment by Urea Wash was elucidated and compared to Alkaline Wash and without washing.