Marshall T, Williams K M
Biochemistry Research Laboratory, School of Pharmaceutical & Chemical Sciences, Sunderland Polytechnic, Green Terrace, Great Britain.
Electrophoresis. 1991 Jul-Aug;12(7-8):461-71. doi: 10.1002/elps.1150120703.
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.
本文综述了我们简化的高分辨率二维聚丙烯酰胺凝胶电泳(2-D PAGE)技术在人体体液中的应用。展示了与酒精滥用、家族性异常脂蛋白血症(“鱼眼病”)和心肌梗死相关的血清/血浆蛋白变化。参照他人的研究成果,展示了羊水、脑脊液、尿液和唾液的高分辨率2-D PAGE,并首次报道了在汗液和泪液中检测到粉紫色染色的“荧光胺染料”。讨论了该方法的一般方面。这些方面包括样品制备、电泳条件(变性或非变性)的选择和检测方法(考马斯亮蓝或银染),以及在银染前用十二烷基硫酸钠对天然蛋白质进行预处理或在二维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳前在甘油中进行等电聚焦凝胶收缩的影响。