The simplified technique of high resolution two-dimensional polyacrylamide gel electrophoresis: biomedical applications in health and disease.
The application of our simplified technique of high resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) to human body fluids is reviewed. Serum/plasma protein changes associated with alcohol abuse, familial dyslipoproteinemia ("fish-eye" disease), and myocardial infarction are demonstrated. High resolution 2-D PAGE of amniotic fluid, cerebrospinal fluid, urine, and saliva is shown with reference to the work of others, and the detection of pink-violet staining "lumicarmines" in sweat and tear fluid is reported for the first time. General aspects relating to the methodology are discussed. These include sample preparation, the choice of electrophoresis conditions (denaturing or nondenaturing) and detection method (Coomassie Brilliant Blue or silver), and the effects of native protein pretreatment with sodium dodecyl sulfate prior to silver staining or isoelectric focusing gel shrinkage in glycerol prior to second-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis.
Biochemistry Research Laboratory, School of Pharmaceutical & Chemical Sciences, Sunderland Polytechnic, Green Terrace, Great Britain.
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Marshall
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Williams
D000437:Alcoholism / Q000097:blood; D001798:Blood Proteins / Q000032:analysis*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods; D006801:Humans; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods; D008970:Molecular Weight; D009203:Myocardial Infarction / Q000097:blood; D011506:Proteins / Q000032:analysis*; D011507:Proteinuria; D012016:Reference Values; D012471:Salivary Proteins and Peptides / Q000032:analysis; D013542:Sweat / Q000737:chemistry; D013666:Tears / Q000737:chemistry
D001798:Blood Proteins; D011506:Proteins; D012471:Salivary Proteins and Peptides