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Application of binary buffer systems to free flow cell electrophoresis
Author(s) -
Weber Gerhard,
Grimm Daniela,
Bauer Johann
Publication year - 2000
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/(sici)1522-2683(20000101)21:2<325::aid-elps325>3.0.co;2-p
Subject(s) - electrophoresis , isotachophoresis , chromatography , buffer (optical fiber) , chemistry , free flow electrophoresis , capillary electrophoresis , polyacrylamide gel electrophoresis , biochemistry , gel electrophoresis of proteins , electrode , telecommunications , computer science , electrolyte , enzyme
The applicability of free flow electrophoresis (FFE) was expanded towards processing of sensitive cells. The chamber medium was adjusted to a physiologic pH of 7.35 by a mixture of N ‐(2‐hydroxyethyl)piperazine‐ N′ ‐(3‐propanesulfonic acid) (EPPS) and 2,2‐bis(hydroxymethyl)‐2,2′2″‐nitrilotriethanol (BISTRIS). These substances proved to be nontoxic to sensitive cells such as human smooth muscle or thyroid cells. They enhanced the electrical conductivity of the medium only slightly so that a new cell electrophoresis separation medium could be prepared, which contained 30 m M NaCl together with or without 1 m M CaCl 2 but did not generate problems of overheating the fluid. Suspended in this medium, human smooth muscle cells as well as human thyroid carcinoma cells remained viable single cells for at least 120 min. After this period they could be recultured to form monolayers. If electrophoresed in the Octopus preparative FFE device, they migrated as single cells and did not clot; therefore, their electrophoretic behavior could be determined exactly.