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Separation and isolation of subcellular‐size particles by electrophoresis in polymer solution using the commercial scanning apparatus
Author(s) -
Chang HuangTung,
Zakharov Sergey F.,
Chrambach Andreas
Publication year - 1996
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.1150170426
Subject(s) - polyvinylpyrrolidone , chromatography , electrophoresis , polystyrene , fluorescence , polymer , scanning electron microscope , fluorescein , analytical chemistry (journal) , chemistry , materials science , polymer chemistry , optics , physics , organic chemistry , composite material
Electrophoresis of fluorescently labeled rat liver microsomes and polystyrene carboxylates of 10 and 30 nm radius was conducted in buffered 10–15% polyvinylpyrrolidone ( M r = 10 6 ) solutions, using a horizontal gel electrophoresis apparatus with intermittent scanning of fluorescence (HPGE‐1000, LabIntelligence). Banding, constant migration rates and Ferguson plots were obtained in these polymer solutions. The major microsome band detected by the automated scan was located visually on the gel by means of its fluorescein label and was isolated by volumetric withdrawal; recovery was monitored by scanning and ascertained to be near quantitative after three consecutive steps, in each of which 30 μL were withdrawn. This preparative method promises to be generally applicable to particles that are too large to enter into gels.