Separation and detection of selenium-containing proteins in human serum.
Selenium-containing proteins or their subunits in human serum were separated and detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and the amount of selenium in each protein band was determined by HPLC with a fluorescence detector after derivatization with 2,4-diaminonaphthalene (DAN). This procedure provides a detection limit of 0.06 ng in a linear range of 0-1.5 ng. A protein is defined as a selenium-containing protein if its mean Se content exceeds twice the detection limit (0.12 ng) and twice the standard deviation of three replicates in sample determination. At least 4 selenium-containing bands with apparent molecular masses of 57-74, 46-56, 40-42 and 21-22 kDa could be detected from human serum collected from 4 volunteers.
State Key Laboratory of Environmental Aquatic Chemistry, Research Center for Eco-Environmental Sciences, Beijing. wanzj@mail.rcees.ac.cn
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Gao
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Wang
D015081:2-Naphthylamine / Q000031:analogs & derivatives; D000328:Adult; D001798:Blood Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005433:Flour; D005453:Fluorescence; D006801:Humans; D008970:Molecular Weight; D012275:Oryza / Q000737:chemistry; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012643:Selenium / Q000032:analysis*; D051140:Selenoproteins; D012680:Sensitivity and Specificity
D001798:Blood Proteins; D011506:Proteins; D051140:Selenoproteins; D015081:2-Naphthylamine; D012643:Selenium