Urolithiasis 2 pp 285-286 | Cite as

Two-Dimensional Gel Analysis of Proteins in Unprocessed Human Urine Using Double Stain

  • P. K. Grover
  • M. I. Resnick

Abstract

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.

Keywords

Albumin Urea Electrophoresis Polyacrylamide Oxalate 

References

  1. 1.
    S A Lewis and C Clausen, Urinary proteases degrade epithelial sodium channels, J Membrane Biol 122: 77 (1991).CrossRefGoogle Scholar
  2. 2.
    N Frearson, RD Taylor and SV Perry, Proteins in the urine associated with Duchenne muscular dystrophy and other neuromuscular diseases, Clin Sci 61: 141 (1981).PubMedGoogle Scholar
  3. 3.
    EL Kanabrocki, JA Kanabrocki, RB Sothem et al, Circadian distribution of proteins in urine from healthy young men, Chronobiol Int 7: 433 (1990).PubMedCrossRefGoogle Scholar
  4. 4.
    BM Fraij, Separation and identification of urinary proteins and stone-matrix proteins by mini-slab sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Clin Chem 35: 658 (1989).PubMedGoogle Scholar
  5. 5.
    J Guevara, DA Johnston, LS Ramagali et al, Quantitative aspects of silver deposition in proteins resolved in complex polyacrylamide gels, Electrophoresis 3: 197 (1982).CrossRefGoogle Scholar
  6. 6.
    B Hess, Y Nakagawa, JH Parks and FL Coe, Molecular abnormality of Tamm-Horsfall glycoprotein in calcium oxalate nephrolithiasis, Amer J Physiol 260: F569 (1991).PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • P. K. Grover
    • 1
  • M. I. Resnick
    • 1
  1. 1.Division of UrologyUniversity Hospitals of ClevelandClevelandUSA

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