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Size-exclusion high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins: A comparison

✍ Scribed by Dj. Josić; H. Baumann; W. Reutter

Publisher
Elsevier Science
Year
1984
Tongue
English
Weight
933 KB
Volume
142
Category
Article
ISSN
0003-2697

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✦ Synopsis

When 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 a-subunit of the acetylcholine receptor was shown under reducing and nonreducing conditions. 0 1984 Academic press. IX.

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