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Isolation of subcellular‐sized particles separated by electrophoresis in dilute polymer solution, using commercial electrophoresis apparatus with intermittent scanning of fluorescence
Author(s) -
Chen g,
Chrambach Andreas
Publication year - 1997
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.1150180520
Subject(s) - electrophoresis , polyvinylpyrrolidone , chromatography , polymer , resolution (logic) , capillary electrophoresis , chemistry , particle (ecology) , fluorescence , analytical chemistry (journal) , materials science , polymer chemistry , organic chemistry , computer science , oceanography , physics , quantum mechanics , artificial intelligence , geology
Abstract Resolution of subcellular‐sized particles in electrophoresis employing semidilute polymer solutions as “sieving media” improves as the polymer concentration is decreased. Therefore, the previously reported conditions of preparative electrophoresis of microsomes, using concentrated (12%) polyvinylpyrrolidone (PVP) solutions, while solving the problem of non‐entrance of large particles into “sieving media”, do not provide adequate resolving capacity, as exemplified by failure of the microsome preparation used, to resolve in the manner of gels or dilute solutions. The present report provides the conditions under which the HPGE‐1000 apparatus can be preparatively applied when the electrophoretic separation is effectively conducted in a dilute polymer solution. The isolation of three microsome components under those conditions constitutes the first application of “particle sieving”, i.e. , a separation due preponderantly to size and shape differences, at a preparative scale.