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  • Title: Two-dimensional gel electrophoresis of rat liver microsomal membrane proteins.
    Author: Kaderbhai MA, Freedman RB.
    Journal: Biochim Biophys Acta; 1980 Sep 02; 601(1):11-21. PubMed ID: 7407157.
    Abstract:
    Total rat liver microsomal proteins are not suitable for isoelectric focusing in polyacrylamide gels, even in the presence of sodium dodecyl sulphate and excess non-ionic detergent; considerable quantities of protein form an aggregate in the isoelectric focusing gel. This prevents resolution of microsomal proteins by the increasingly popular two-dimensional electrophoresis technique employing isoelectric focusing followed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The problem is caused by the extreme insolubility of some microsomal proteins, especially cytochrome P-450 species, which precipitate during isoelectric focusing. A selective extraction of microsomes with sodium deoxycholate excludes these poorly soluble proteins. The extracted proteins can then be resolved without difficulty by isoelectric focusing, and give excellent two-dimensional gel patterns showing more than 100 proteins, mainly in the pI range 5--7. The technique should be useful in studies on microsome protein topology and on changes in microsome composition.
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