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  • 1
    Electronic Resource
    Electronic Resource
    Optimized continuous flow electrophoresis (1996)
    Weinheim : Wiley-Blackwell
    Electrophoresis 17 (1996), S. 1906-1910 
    Details
    ISSN: 0173-0835
    Keywords: Continuous flow electrophoresis ; Free flow electrophoresis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Continuous flow electrophoresis (CFE) was optimized by employing (i) electrophoretic regimes with stacking properties, to eliminate electrohydrodynamic dispersion, (ii) quasi-mixed zones to prevent precipitation of the stacked analytes, (iii) sheath liquid streams at the electrode compartment membranes to prevent penetration of the electrode reaction products into the separation chamber, (iv) proper engineering of the separation chamber to provide efficient dissipation of Joule heat, and (v) counterflow at the collection outlets to eliminate the problems of dead volumes and uneven collection of separated species. Data on direct temperature measurements in the separation chamber at various levels of the dissipated electric power are presented. Preparative runs of amyloglucosidase in the isoelectric focusing (IEF) mode and rat liver organelles in the isotochophoresis (ITP) mode demonstrate the high performance of the optimized CFE system.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150171216
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  • 2
    Electronic Resource
    Electronic Resource
    Recent developments in preparative free flow isoelectric focusing (1998)
    Weinheim : Wiley-Blackwell
    Electrophoresis 19 (1998), S. 1649-1653 
    Details
    ISSN: 0173-0835
    Keywords: Free flow electrophoresis ; Preparative electrophoresis ; Continuous isoelectric focusing ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Continuous flow electrophoresis (CFE) is a promising method for preparative fractionation of a variety of biological species, ranging from peptides and proteins to subcellular particles and cells. The high separation efficiency of FFE may be deteriorated by hydrodynamic distortion of zones due to the omnipresent parabolic laminar flow profile. We show in this paper that the detrimental hydrodynamic distortion of separated proteins zones can be reduced, with resultant enhancement of separation efficiency, by employing continuous isoelectric focusing in pH gradients as the actual working regime in an advanced instrumentation. Newly developed media for fast generation of narrow- or broad-range pH gradients under CFE conditions are described. The separation efficiency of these pH gradients is comparable to that of the gradients formed with the aid of synthetic carrier ampholytes. The new media are defined mixtures of nontoxic chemicals, and thus they are compatible with the requirements of human medicine. Experimental data are given showing that the new media offer fractionation of isoforms of proteins, that they offer resolution of proteins differing in isoelectric point (pI) by less than 0.05 pH units, and that these media inhibit proteins precipitation in experiments with human serum proteins.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150191021
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  • 3
    Electronic Resource
    Electronic Resource
    Sodium chloride in separation medium enhances cell compatibility of free flow electrophoresis (1995)
    Weinheim : Wiley-Blackwell
    Electrophoresis 16 (1995), S. 92-97 
    Details
    ISSN: 0173-0835
    Keywords: Free flow electrophoresis ; Margin buffers ; Sodium chloride ; Mononuclear leukocytes ; Fura2-AM ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: 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 mM 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 mM NaCl were present could an undisturbed cytosolic Ca2+ metabolism be maintained for the time of a free flow electrophoresis cell separation experiment.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150160116
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  • 4
    Electronic Resource
    Electronic Resource
    Sodium chloride in preparative free-flow cell electrophoresis: Recent developments (1996)
    Weinheim : Wiley-Blackwell
    Electrophoresis 17 (1996), S. 526-528 
    Details
    ISSN: 0173-0835
    Keywords: Free flow electrophoresis ; Cell electrophoresis ; Margin buffers ; Sodium chloride ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Three buffer systems for free-flow electrophoresis have been designed, which proved useful for performing cell electrophoresis in the presence of 50 mM NaCl. Each system consists of one central cell suspension buffer with 50 mM NaCl, two margin buffers, and two electrode buffers. With the aid of a bromophenol blue/maxilon blue accumulation test the various buffers were adjusted to ensure a laminar flow and remain unchanged on their way through an electrophoresis chamber.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150170319
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  • 5
    Electronic Resource
    Electronic Resource
    Free flow-isoelectric focusing of human cellular lysates as sample preparation for protein analysisPresented at the Electrophoresis Forum '94, October 24-26, 1994, at the Technical University Munich, Germany (1995)
    Weinheim : Wiley-Blackwell
    Electrophoresis 16 (1995), S. 1010-1015 
    Details
    ISSN: 0173-0835
    Keywords: Free flow electrophoresis ; Isoelectric focusing ; Two-dimensional polyacrylamide gel electrophoresis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A whole cell lysate of human cells was separated into 80 fractions according to the pI of proteins using free flow isoelectric focusing with carrier ampholytes. The resolution of the process was highly reproducible, with an overlap of fractions of less than 30%. A protein of a faint silver stained spot in two-dimensional gel electrophoresis (2-DE) could be enriched, yielding a Coomassie blue stained spot which could be further characterized by proteinchemical methods. The enrichment of less abundant proteins from a complex crude cell extract was found to be a suitable tool for sample preparation and enrichment before applying proteins to 2-DE and reversed-phase high performance liquid chromatography.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.11501601169
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