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Ultrathin‐layer sodium dodecyl sulfate‐polyacrylamide gradient gel electrophoresis and silver staining of urinary proteins
Author(s) -
Schiwara HansWalter,
Hebell Thomas,
Kirchherr Hartmut,
Postel Wilhelm,
Weser Johannes,
Görg Angelika
Publication year - 1986
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150071104
Subject(s) - silver stain , staining , coomassie brilliant blue , chemistry , chromatography , gel electrophoresis , sodium dodecyl sulfate , polyacrylamide gel electrophoresis , albumin , electrophoresis , gel electrophoresis of proteins , sodium , urine , two dimensional gel electrophoresis , biochemistry , microbiology and biotechnology , enzyme , proteomics , biology , gene , genetics , organic chemistry
Ultrathin‐layer sodium dodecyl sulfate‐polyacrylamide gradient gel electrophoresis in combination with silver staining is a highly sensitive and rapid method for molecular size analysis of urinary proteins. First results are achieved after 4 h (electrophoretic run, 100 min, followed by Coomassie Brilliant Blue staining, 2 h). After additional silver staining, including a recycling step, the results are ready for evaluation within 7 h. Urine samples do not need to be concentrated. Within a single gel, 25–27 samples can be run side by side under identical conditions and thus compared without ambiguity. By coelectrophoresis of pure proteins and immunological analysis 13 urinary proteins could be identified and for 7 of them the detection limits were determined. Glomerular protein patterns in samples of urine with a protein concentration in the normal range are compared with the concentration of albumin and transferrin measured by a sensitive enzyme immunoassay. Typical renal and extrarenal protein patterns are shown.