Two-Dimensional Gel Analysis of Proteins in Unprocessed Human Urine Using Double Stain
Analysis of human urine for protein composition is helpful to diagnose various dysfunctions particularly of renal origin. However, in order to detect the excretion of protein(s) in a given disease, it is crucial to identify proteins in urine of healthy subjects which serve as control. Though two-dimensional (2-D) gel electrophoresis was employed in the past to scan urinary proteins, the studies should be regarded with considerable scepticism as there have been serious shortcomings. For instance, despite the inherent protease activity1 the urines were never collected over protease inhibitors. Only individual urines were analysed, though there were reports of biologic and rhythmic variations in the excretion of proteins2, 3. The samples were refrigerated during the collection period and centrifuged and filtered. This processing of human urine has now been reported to remove various proteins4. Also, though silver stain is 370 fold more sensitive than conventional Coomassie Brilliant Blue5, proteins in the gels were visualized often by the latter stain.
KeywordsAlbumin Urea Electrophoresis Polyacrylamide Oxalate