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Analysis of proteins in renal tubule fluid by ultramicrogradient gel electrophoresis in 0.5 microliter capillary tubes
Author(s) -
Manz Nany,
Galaske Rainer G.,
Cho Joan,
Van Liew Judith B.
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.1150070408
Subject(s) - capillary action , chemistry , chromatography , tubule , capillary electrophoresis , renal tubule , acrylamide , reproducibility , electrophoresis , kidney , materials science , polymer , biology , organic chemistry , copolymer , composite material , endocrinology
A research method is described for producing ultramicrogradient acrylamide gels in 0.5 μL constant volume capillary tubes (inner diameter 130 μm and 32 mm length) suitable for the determination of protein in kidney tubule fluid. Previous methods have been limited to gels contained in 1 and 5 μL capillary tubes. The acrylamide gel concentration varies from 4 to 27% in a functional gel of approximately 40 nL and a length of 2.0 to 2.9 mm. Relative migration distances in these gels of known proteins of varying molecular weight (6000 to 669 000) are comparable to those measured in gels contained in 5.0 μL capillary tubes. After electrophoresis, the gels are extruded from the capillary tubes, stained with Fast Green, destained and scanned in a Joyce‐Loebl microdensitometer with integrator unit. Such gels are capable of retaining and quantitating low molecular weight proteins contained in nanoliter volumes of biological fluids. The reproducibility of this technique for 0.5 μL capillary gels is comparable to that achieved with the routine analysis of proteins in unconcentrated urine using 5.0 μL gels. Examples are given of protein analyses in kidney tubule fluids and urine obtained from normal rats and those with glomerulonephritis.