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Sodium chloride in separation medium enhances cell compatibility of free flow electrophoresis
Author(s) -
Bondy Brigitta,
Bauer Johann,
Seuffert Ingrid,
Weber Gerhard
Publication year - 1995
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.1150160116
Subject(s) - electrophoresis , chromatography , free flow electrophoresis , buffer (optical fiber) , chemistry , sodium , capillary electrophoresis , sodium citrate , analytical chemistry (journal) , biochemistry , polyacrylamide gel electrophoresis , gel electrophoresis of proteins , medicine , telecommunications , organic chemistry , pathology , computer science , enzyme
Free flow electrophoresis of cell suspensions in buffers containing sodium chloride was investigated using a modified procedure and the new apparatus Octopus PZE. The major methodical innovations are upward fluid flow margin buffers flowing through the electrophoresis chamber at both sides of a central cell suspension buffer, adjacent to the electrode membranes, and a sample injection device which focuses the cells hydrodynamically to the middle of the chamber thickness. Mononuclear leukocytes, suspended in a buffer containing 35 m M NaCl, could be fractionated with the same accuracy as by conventional free flow electrophoresis, operated with a single NaCl‐free chamber buffer. However, testing the vitality of separated cells with the help of the calcium indicator FURA2‐AM clearly demonstrated the biological importance of the presence of a minimum amount of sodium chloride during cell electrophoresis. Only if at least 35 m M NaCl were present could an undisturbed cytosolic Ca 2+ metabolism be maintained for the time of a free flow electrophoresis cell separation experiment.