Size-exclusion high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins: A comparison
DOI: 10.1016/0003-2697(84)90492-5
Title: Size-exclusion high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins: A comparison
Journal Title: Analytical Biochemistry
Volume: 142
Issue: 2
Publication Date: 1 November 1984
Start Page: 473
End Page: 479
Published online: online 6 December 2004
ISSN: 0003-2697
Affiliations:

  • a Institut für Molekularbiologie und Biochemie der Freien Universität Berlin, Arnimallee 22, 1000 Berlin 33 (Dahlem), West Germany

  • Institut für Biochemie der Freien Universität Berlin, Fabeckstr. 34-36, 1000 Berlin 33 (Dahlem), West Germany
  • Abstract: rong>Size-exclusion HPLC (SE-HPLC) is applied for the separation of hydrophilic and hydrophobic proteins, numerous problems can be encountered, which may present considerable difficulties. A major source of such complications is interaction between column packing and sample, especially on the so-called “Diol” and “Polyol” columns. In many cases interaction can be reduced only by adding detergents. Calibration proteins and hydrophobic membrane proteins of liver are separated by SE-HPLC. The influence of detergents on association and dissociation of protein subunits and protein configuration is shown. These factors can affect the elution volume during chromatography. Furthermore it is shown that a direct comparison can be drawn between the protein separation by SE-HPLC on the one hand and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis on the other hand. As an example the separation of the δ-subunit of the acetylcholine receptor was shown under reducing and nonreducing conditions.
    Received: 28 February 1984
    Keywords: SE-HPLC of hydrophobic and hydrophilic proteins; influence of detergents; comparison between separation by SE-HPLC and SDS-PAGE

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