
原发性肾病综合征儿童尿液双向电泳图谱的建立
Establishment of urinary proteomic map on two-dimensional polyacrylamide gel electrophoresis in children with idiopathic nephrotic syndrome
目的:建立与优化原发性肾病综合征(INS)儿童尿液全蛋白双向凝胶电泳技术(2-DE)。方法:采用4种不同的方法制备蛋白质样本, 以固相pH梯度等电聚焦为第一向, 12%的SDS-PAGE进行第二向电泳,银染显色。结果:以丙酮浓缩蛋白后透析提纯法,制备蛋白质标本,300 μg为上样量时,可获得图像清晰、分辨率高、重复性好的儿童INS尿液2-DE图谱。结论:建立了稳定的原发性肾病综合征儿童尿液双向凝胶电泳技术,为进一步开展疾病的尿液蛋白质组学研究奠定了基础。[中国当代儿科杂志,2007,9(2):122-124]
OBJECTIVE: To establish a urinary proteomic map on two-dimensional polyacrylamide gelelectrophoresis (2-DE) in children with idiopathic nephrotic syndrome (INS). METHODS: The proteins from INS children were purified by four various means, separated by 2-DE, and stained by silver. RESULTS The sequential preparation of urinary proteins by acetone precipitation and dislysis, when the sample was 300 μg, resulted in a clear background, well-resolved and reproducible 2-DE urinary protemic map in children with INS. CONCLUSIONS A steady 2-DE technique for urinary protemic map in children with INS was established, which can be effectively applied in urinary proteomics of the disease.[Chin J Contemp Pediatr, 2007, 9 (2):122-124]
Nephrotic syndrome / Urine / Two-dimensional polyacrylamide gel electrophoresis / Child