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Separation of viable from radiation‐induced apoptotic lymphocytes by free‐flow electrophoresis
Author(s) -
Chestkov Valery,
Baibakov Boris,
Radko Sergey P.,
Chrambach Andreas
Publication year - 1998
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.1150190723
Subject(s) - electrophoresis , apoptosis , chemistry , chromatography , free flow electrophoresis , population , in vitro , flow cytometry , lymphocyte , microbiology and biotechnology , triethanolamine , biophysics , biochemistry , biology , analytical chemistry (journal) , polyacrylamide gel electrophoresis , immunology , gel electrophoresis of proteins , demography , sociology , enzyme
A human lymphocyte population undergoing apoptosis in vitro due to γ‐irradiation was fractionated by free‐flow electrophoresis in triethanolamine – Na‐acetate buffers, containing up to 50 m M NaCl, with pH 6.0, 7.2 and 8.5, made isotonic by addition of sucrose. As shown by a flow cytometric analysis of the eluate, the distribution of apoptotic lymphocytes is shifted to the range of higher electrophoretic mobilities relative to that of viable ones at pH 8.5, yielding cell fractions enriched in apoptotic cells by a factor of 3 to 5. The difference in rates of electrophoretic migration observed at a mildly alkaline pH but not at a neutral or mildly acidic one suggests that the surface of apoptotic lymphocytes is more acidic than that of viable ones.