The determination of specific radioactivity of proteins eluted intact from polyacrylamide gels, utilizing a fluorescamine assay.
The methodology described permits the measurement of the specific radioactivity of diverse proteins resolvable by separatory techniques using cylindrical polyacrylamide gels. Following separation, the proteins are electroeluted; eluted protein is quantitated in the microgram range using a fluorescamine assay, while the major portion of the recovered sample is used for radioactivity measurement. These procedures have been adapted for use in tracer studies of protein metabolism. Their utility in kinetic investigations is demonstrated with data on the time course of changing specific radioactivities of human plasma albumin and apolipoprotein B labeled in vivo with a [3H]leucine tracer.
D001055:Apolipoproteins B / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005450:Fluorescamine; D006801:Humans; D007457:Iodine Radioisotopes; D011506:Proteins / Q000032:analysis*; D012709:Serum Albumin / Q000032:analysis; D012711:Serum Albumin, Radio-Iodinated; D013050:Spectrometry, Fluorescence; D013141:Spiro Compounds; D014316:Tritium
D001055:Apolipoproteins B; D007457:Iodine Radioisotopes; D011506:Proteins; D012709:Serum Albumin; D012711:Serum Albumin, Radio-Iodinated; D013141:Spiro Compounds; D014316:Tritium; D005450:Fluorescamine