A rapid flat gel isoelectric focusing method for the determination of apolipoprotein E phenotypes and its application.
A rapid flat gel isoelectric focusing method has been developed for the determination of apolipoprotein E phenotypes. Isoelectric focusing in 5% polyacrylamide flat gel with 8 mol/l urea and 2.8% pharmalyte (pH 4-6.5) was carried out at 3,000 V and 4 degrees C for 1 h under a constant power of 30 W. The separation of apolipoprotein E isoproteins was good and the isoelectric points were determined. Using this method, the apolipoprotein E phenotype frequency was examined in the Japanese population, and a higher frequency of phenotype E3/3 and a lower frequency of phenotype E3/2 were found in Japanese than those reported for the German, American or English population. In our focusing system the cut-off point of apolipoprotein E2/E3 ratio between the phenotype E3/3 and E3/2 was assumed to be approximately 0.9. These results indicate that this method may be useful for the determination of apolipoprotein E phenotypes.
D000180:Acrylic Resins; D000293:Adolescent; D000328:Adult; D000368:Aged; D001057:Apolipoproteins E / Q000235:genetics* / Q000302:isolation & purification; D005260:Female; D005787:Gene Frequency; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D007564:Japan; D008297:Male; D008875:Middle Aged; D010641:Phenotype
D000180:Acrylic Resins; D001057:Apolipoproteins E; C016680:polyacrylamide gels