The use of a hollow fiber membrane module in sample conditioning prior to electrophoresis.
A new method to continuously change the buffer conditions in samples prior to electrophoresis by counter-current dialysis has been developed using a hollow fiber membrane module (VariPerm M, bitop, Witten). It is demonstrated that the collected fractions of a column eluate after hydrophobic interaction chromatography cannot be analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) due to the high content of ammonium sulfate. In contrast, when the hollow fiber membrane module is used for on-line desalting of the eluate during chromatography, it is possible to detect distinct bands on SDS-gels even after sample concentration by evaporation. Moreover, adding polyethyleneglycol 20,000 to the counter-current dialysis buffer the hollow fiber membrane module is suitable for combined on-line sample desalting and sample concentration of living organisms during free flow electrophoresis (FFE): a culture of Nitrosomonas eutropha which could not significantly be detected by FFE without conditioning was successfully analyzed when concentrated and desalted on-line during sample application.
Institut für Biochemie, Universität Witten/Herdecke, Germany.
T
T
Schwarz
S
S
Sindern
P
P
Bartholmes
M
M
Kaufmann
D000645:Ammonium Sulfate; D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification*; D002197:Capillary Action; D003956:Dialysis / Q000295:instrumentation / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation* / Q000379:methods; D005056:Euglena gracilis / Q000737:chemistry; D007202:Indicators and Reagents; D009606:Nitrosomonas / Q000737:chemistry; D011092:Polyethylene Glycols; D015800:Protozoan Proteins / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate
D001426:Bacterial Proteins; D007202:Indicators and Reagents; D015800:Protozoan Proteins; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D000645:Ammonium Sulfate