血清双向凝胶电泳体系的建立

目的:建立血清双向凝胶电泳(two-dimensional polyacrylamide gel electrophoresis,2-DE)技术体系.方法:用Proteoprep blue albumin depletion kit将血清中高峰度蛋白去除.预冷丙酮沉淀蛋白质后,加入样品溶解液充分溶解,进行等电聚焦电泳(isoelectric focusing,IEF)和十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate-polyacrylamide gel elecrophoresis,SDS-PAGE),行硝酸银染色后,运用ImageMaster软件行初步分析.结果:通过调整,优化2-DE技术条件,建立了稳定的2-DE技术.结论:建立了相对稳定的人血清蛋白质2-DE技术,其稳定性、重复性好,为开展疾病蛋白质组学的研究奠定了基础.

Establishment of Two-dimensional Polyacrylamide Gel Electrophoresis System for Serum Study

Objective:To establish a two-dimensional polyacrylamide gel electrophoresis(2-DE) technique for serum study.Methods: After elimination of serum albumin and IgG was achieved by filtering with proteoprep blue albumin depletion kit,proteins of interest were precipitated with acetone pre-cooling and subjected to adequate dissolution.Isoelectric focusing electrophoresis(IEF) was employed as the first dimension and sodium dodecyl sulfate-polyacrylamide gel elecrophoresis(SDS-PAGE) the second dimension.After silver nitrate staining,protein spots were analyzed.Results:A steady 2-DE technique procedure was established under opitimized conditions.Conclusion:The established 2-DE technique of human serum proteins can be effecting applied in serum proteomics of diseases.